In this study, we investigated daily blood sugar (BS)

changes in NAFLD patients using CGMS. Sixty-five patients; 35 female, median age 61 years, median body mass index (BMI) 27.1 with biopsy-proven NAFLD according to Brunt’s fibrosis stage; 9 patients of F1, 23 of F2, 18 of F3, and 15 of F4, were enrolled. We performed 75g oral glucose tolerance tests (OGTT) in 28 patients with <140mg/dl fasting BS without a diagnosis of DM before enrollment, and the changes in BS during 24 hours by Medtronic iPro2® CGMS were evaluated find more in all 65 patients. Of 37 patients with DM including 3 diagnosed by OGTT, 7 received insulin injections, 3 sulfonylurea (SU), 3 metformin (Met), 8 DPP4 antagonist (DPP4), 5 Met +DPP4, 3 SU+DPP4, 3 SU+Met+DPP4, and 12 dietary therapy alone. Informed consent in writing was obtained from each patient and the study protocol conformed

to the ethical guidelines of the 1975 Declaration of Helsinki and our institutional review committee. The prevalence of DM was significantly higher with the progression of hepatic fibrosis, at 80% in patients with cirrhosis vs. 50% without cirrhosis. CGMS revealed variability of median BS, standard deviation of median BS, maximum (max) BS, and differences in max and minimum (min) BS to be significantly Atezolizumab price higher in cirrhotic patients (0.01, 0.01, 0.02, and 0.003, respectively). Postprandial hyperglycemia exceeding 300 mg/dl and a difference between max and min BS over 200

mg/dl were seen only in 5 cirrhotic patients with DM. Interestingly, nocturnal hypoglycemia with BS<60mg/dl was seen in 7 males with remarkably high serum insulin levels (median selleck chemical serum fasting immunoreactive insulin level 27.6 μU/ mL); median age 31 years, 6 patients with super-obesity; BMI >35, 4 diagnosed with impaired glucose tolerance, 6 in F1 or F2, and none being treated with anti-diabetic drugs. CGMS analysis revealed postprandial hyperglycemia in cirrhotic patients and nocturnal hypoglycemia in relatively young and highly overweight males with severe IR and mild fibrosis revealed to be characteristic of NAFLD patients. The latter might predict both the progression of hepatic fibrosis and a poor outcome. Disclosures: The following people have nothing to disclose: Makiko Taniai, Etsuko Hashimoto, Kazuhisa Kodama, Tomomi Kogiso, Katsutoshi Tokushige, Keiko Shiratori Objectives: Diabetes and fatty liver (FL) disease are risk factors for hepatocellular carcinoma and cardiovascular disease. However, the effect of fatty liver in diabetes remains unclear. We tried to elucidate the roles of fatty liver in diabetes related to prognosis, including HCC, extrahepatic tumor, and cardiovascular events. Methods: Study design: Prospective cohort study.

In this study, we investigated daily blood sugar (BS)

changes in NAFLD patients using CGMS. Sixty-five patients; 35 female, median age 61 years, median body mass index (BMI) 27.1 with biopsy-proven NAFLD according to Brunt’s fibrosis stage; 9 patients of F1, 23 of F2, 18 of F3, and 15 of F4, were enrolled. We performed 75g oral glucose tolerance tests (OGTT) in 28 patients with <140mg/dl fasting BS without a diagnosis of DM before enrollment, and the changes in BS during 24 hours by Medtronic iPro2® CGMS were evaluated Cobimetinib supplier in all 65 patients. Of 37 patients with DM including 3 diagnosed by OGTT, 7 received insulin injections, 3 sulfonylurea (SU), 3 metformin (Met), 8 DPP4 antagonist (DPP4), 5 Met +DPP4, 3 SU+DPP4, 3 SU+Met+DPP4, and 12 dietary therapy alone. Informed consent in writing was obtained from each patient and the study protocol conformed

to the ethical guidelines of the 1975 Declaration of Helsinki and our institutional review committee. The prevalence of DM was significantly higher with the progression of hepatic fibrosis, at 80% in patients with cirrhosis vs. 50% without cirrhosis. CGMS revealed variability of median BS, standard deviation of median BS, maximum (max) BS, and differences in max and minimum (min) BS to be significantly selleckchem higher in cirrhotic patients (0.01, 0.01, 0.02, and 0.003, respectively). Postprandial hyperglycemia exceeding 300 mg/dl and a difference between max and min BS over 200

mg/dl were seen only in 5 cirrhotic patients with DM. Interestingly, nocturnal hypoglycemia with BS<60mg/dl was seen in 7 males with remarkably high serum insulin levels (median selleck chemicals serum fasting immunoreactive insulin level 27.6 μU/ mL); median age 31 years, 6 patients with super-obesity; BMI >35, 4 diagnosed with impaired glucose tolerance, 6 in F1 or F2, and none being treated with anti-diabetic drugs. CGMS analysis revealed postprandial hyperglycemia in cirrhotic patients and nocturnal hypoglycemia in relatively young and highly overweight males with severe IR and mild fibrosis revealed to be characteristic of NAFLD patients. The latter might predict both the progression of hepatic fibrosis and a poor outcome. Disclosures: The following people have nothing to disclose: Makiko Taniai, Etsuko Hashimoto, Kazuhisa Kodama, Tomomi Kogiso, Katsutoshi Tokushige, Keiko Shiratori Objectives: Diabetes and fatty liver (FL) disease are risk factors for hepatocellular carcinoma and cardiovascular disease. However, the effect of fatty liver in diabetes remains unclear. We tried to elucidate the roles of fatty liver in diabetes related to prognosis, including HCC, extrahepatic tumor, and cardiovascular events. Methods: Study design: Prospective cohort study.

Latitude and longitude were correlated, so to avoid multicollinearity we used the first principal component of these two variables as our geographic factor. This principal component explained 84% of the variance in latitude and longitude. We used the same model to test for elevation effects on each of the three song features by removing the geographic

factor and replacing it with elevation values for the site of each recording. We ran these three tests separately from the previous three because the geographic factor was correlated with elevation. All statistics were performed in jmp version 9.0 (SAS Institute Inc., Cary, NC, USA, Gemcitabine purchase 2010). All measured songs were discrete and brief with breaks between songs lasting at least 1 s. All birds sang songs that included one or two introductory

note types, followed by a more complex end phrase that often ran into a trill (Figs 2-4). selleck inhibitor Songs with two different introductory note types were rare, and 87% of songs included only two syllable types: an introductory note repeated up to six times, followed by an end phrase that was sometimes repeated more than once. We observed three general introductory note types: (1) a descending frequency sweep (46 birds); (2) a broadband buzz (32 birds); (3) a harmonic stack (5 birds) (Fig. 2). Nineteen birds sang multiple introductory note types, with one singing all three introductory notes. Acoustically, these notes were not all identical among individuals, but were clearly identifiable to type. Song end phrases showed much more diversity in form than introductory notes (Fig. 3). We identified 77 end-phrase types. Of these, 42 were unique to single birds; the remaining 35 end phrases were shared by two to seven individuals each. Frequency characteristics of the three introductory note types, end phrases and whole songs are detailed in Table 1. In total, we identified 179 song types from the 61 recorded birds. The largest recorded repertoire

of an individual included 16 song types and 15 syllable types. Individuals typically sang with eventual variety. Longer recordings contained selleck kinase inhibitor more song types (Appendix S1), and all recorded birds continued to produce new song and end-phrase types up to the end of each recording. Thus, we expect that more extensive sampling would discover more song types from every individual. Song bouts may include a wide variety of song forms generated by altering (1) the type of introductory note; (2) the number of repetitions of the introductory note; (3) the type of end phrase; (4) the number of repetitions of the end phrase; (5) the addition of a third note type (Fig. 4). On a local scale, songs were highly varied: multiple tracks (n = 2–4) from nine common locations did not show evidence of song sharing by neighbours.

Latitude and longitude were correlated, so to avoid multicollinearity we used the first principal component of these two variables as our geographic factor. This principal component explained 84% of the variance in latitude and longitude. We used the same model to test for elevation effects on each of the three song features by removing the geographic

factor and replacing it with elevation values for the site of each recording. We ran these three tests separately from the previous three because the geographic factor was correlated with elevation. All statistics were performed in jmp version 9.0 (SAS Institute Inc., Cary, NC, USA, Kinase Inhibitor Library concentration 2010). All measured songs were discrete and brief with breaks between songs lasting at least 1 s. All birds sang songs that included one or two introductory

note types, followed by a more complex end phrase that often ran into a trill (Figs 2-4). Liproxstatin-1 manufacturer Songs with two different introductory note types were rare, and 87% of songs included only two syllable types: an introductory note repeated up to six times, followed by an end phrase that was sometimes repeated more than once. We observed three general introductory note types: (1) a descending frequency sweep (46 birds); (2) a broadband buzz (32 birds); (3) a harmonic stack (5 birds) (Fig. 2). Nineteen birds sang multiple introductory note types, with one singing all three introductory notes. Acoustically, these notes were not all identical among individuals, but were clearly identifiable to type. Song end phrases showed much more diversity in form than introductory notes (Fig. 3). We identified 77 end-phrase types. Of these, 42 were unique to single birds; the remaining 35 end phrases were shared by two to seven individuals each. Frequency characteristics of the three introductory note types, end phrases and whole songs are detailed in Table 1. In total, we identified 179 song types from the 61 recorded birds. The largest recorded repertoire

of an individual included 16 song types and 15 syllable types. Individuals typically sang with eventual variety. Longer recordings contained selleck products more song types (Appendix S1), and all recorded birds continued to produce new song and end-phrase types up to the end of each recording. Thus, we expect that more extensive sampling would discover more song types from every individual. Song bouts may include a wide variety of song forms generated by altering (1) the type of introductory note; (2) the number of repetitions of the introductory note; (3) the type of end phrase; (4) the number of repetitions of the end phrase; (5) the addition of a third note type (Fig. 4). On a local scale, songs were highly varied: multiple tracks (n = 2–4) from nine common locations did not show evidence of song sharing by neighbours.

) Regarding antihypertensive

therapy, a higher proportion of cancer cases had ever used beta-blockers and calcium channel blockers. In the univariate analyses, we found a negative association between pioglitazone/rosiglitazone Ruxolitinib and liver cancer incidence, and a positive one between rosiglitazone and lung cancer. After controlling for potential confounding variables including short-acting human insulin, metformin (mean daily dosage in quartiles), sulfonylurea (mean daily dosage in quartiles), number of oral antidiabetic agents, chronic liver disease, statins, aspirin, beta-blockers, chronic kidney disease, glinides, nephropathy, cerebrovascular disease, calcium channel blockers, cardiovascular disease, and chronic lung disease, a significantly decreased risk of liver cancer incidence was found for any use of rosiglitazone (OR: 0.73, 95% CI: 0.65-0.81) and pioglitazone (OR: 0.83, 95% CI: 0.72-0.95), respectively (Table 3), in contrast to the adjusted ORs of 2.35 (95% CI: Sorafenib chemical structure 2.21-2.49) for short-acting insulin, 1.05 (95% CI: 0.93-1.18) for sulfonylurea, and 0.77 (95% CI: 0.69-0.85) for metformin. The protective effects were even stronger for higher cumulative dosage ≥120 DDD (OR 0.64;

95% CI: 0.56-0.72 for rosiglitazone and OR 0.80; 95% CI: 0.67-0.95 for pioglitazone) and for cumulative treatment duration ≥3 years (OR 0.64; 95% CI: 0.49-0.85 for rosiglitazone and OR 0.44; 95% CI: 0.23-0.86 for pioglitazone). Risk estimates were similar between prevalent and newly diagnosed type 2 diabetes patients. Due to the high prevalence of hepatitis

B and C infection, the analysis was further stratified to those with and without chronic liver disease. The risk reduction (especially in high and prolonged dosage of rosiglitazone and pioglitazone) in liver cancer was mostly seen in the patients with prevalent chronic liver disease (Table 4). For colorectal cancer, rosiglitazone was associated with a significantly decreased risk (OR: 0.86; 95% CI: 0.76-0.96) learn more with a more prominent effect among those with the highest cumulative dose (OR: 0.83; 95% CI: 0.73-0.95) (Table 5). In contrast, pioglitazone was not associated with a significantly protective effect for colorectal cancer, although a trend of negative association was also found. The characteristics for cases and controls of lung and bladder cancer are summarized in Supporting Tables B and C. No relation between rosiglitazone/pioglitazone and lung or bladder cancer was found, although the ORs were above 1.0 (Supporting Tables D, E). A dosage and duration response relationship was not evident between the use of rosiglitazone and pioglitazone and these two types of cancer. An increase in bladder cancer risk was observed with pioglitazone use ≥3 years (OR: 1.56, 95% CI: 0.51-4.74), which did not reach statistical significance. The cancer risk associated with antidiabetic therapies other than TZD are also reported (Supporting Table F).

) Regarding antihypertensive

therapy, a higher proportion of cancer cases had ever used beta-blockers and calcium channel blockers. In the univariate analyses, we found a negative association between pioglitazone/rosiglitazone learn more and liver cancer incidence, and a positive one between rosiglitazone and lung cancer. After controlling for potential confounding variables including short-acting human insulin, metformin (mean daily dosage in quartiles), sulfonylurea (mean daily dosage in quartiles), number of oral antidiabetic agents, chronic liver disease, statins, aspirin, beta-blockers, chronic kidney disease, glinides, nephropathy, cerebrovascular disease, calcium channel blockers, cardiovascular disease, and chronic lung disease, a significantly decreased risk of liver cancer incidence was found for any use of rosiglitazone (OR: 0.73, 95% CI: 0.65-0.81) and pioglitazone (OR: 0.83, 95% CI: 0.72-0.95), respectively (Table 3), in contrast to the adjusted ORs of 2.35 (95% CI: MK-1775 price 2.21-2.49) for short-acting insulin, 1.05 (95% CI: 0.93-1.18) for sulfonylurea, and 0.77 (95% CI: 0.69-0.85) for metformin. The protective effects were even stronger for higher cumulative dosage ≥120 DDD (OR 0.64;

95% CI: 0.56-0.72 for rosiglitazone and OR 0.80; 95% CI: 0.67-0.95 for pioglitazone) and for cumulative treatment duration ≥3 years (OR 0.64; 95% CI: 0.49-0.85 for rosiglitazone and OR 0.44; 95% CI: 0.23-0.86 for pioglitazone). Risk estimates were similar between prevalent and newly diagnosed type 2 diabetes patients. Due to the high prevalence of hepatitis

B and C infection, the analysis was further stratified to those with and without chronic liver disease. The risk reduction (especially in high and prolonged dosage of rosiglitazone and pioglitazone) in liver cancer was mostly seen in the patients with prevalent chronic liver disease (Table 4). For colorectal cancer, rosiglitazone was associated with a significantly decreased risk (OR: 0.86; 95% CI: 0.76-0.96) selleck with a more prominent effect among those with the highest cumulative dose (OR: 0.83; 95% CI: 0.73-0.95) (Table 5). In contrast, pioglitazone was not associated with a significantly protective effect for colorectal cancer, although a trend of negative association was also found. The characteristics for cases and controls of lung and bladder cancer are summarized in Supporting Tables B and C. No relation between rosiglitazone/pioglitazone and lung or bladder cancer was found, although the ORs were above 1.0 (Supporting Tables D, E). A dosage and duration response relationship was not evident between the use of rosiglitazone and pioglitazone and these two types of cancer. An increase in bladder cancer risk was observed with pioglitazone use ≥3 years (OR: 1.56, 95% CI: 0.51-4.74), which did not reach statistical significance. The cancer risk associated with antidiabetic therapies other than TZD are also reported (Supporting Table F).

To our knowledge, this is also the first report showing that the inhibition of miR-152 results functionally in global DNA hypermethylation and increased methylation levels of the TSGs GSTP1 and CDH1 in HCC cell lines. The overexpression of miR-152 in HepG2.2.15 cells reduced GDM from 6.31% to 4.08%, whereas the miR-152 inhibitor in HepG2 cells increased GDM from 4.55% to 5.88%. The GSTP1 gene has been reported to be commonly epigenetically silenced by methylation in HBV-associated HCC, and somatic GSTP1 inactivation may contribute to the pathogenesis of this malignancy.35 In our study, the GSTP1 gene was demonstrated

to be methylated in HepG2 cells, and the methylation level of its promoter that we detected was increased from 58.18% to 86.36% after transfection of the miR-152

inhibitor. CDH1 is also frequently silenced by methylation in HCC, and it has been reported that HBx can repress PD-0332991 nmr CDH1 expression by inducing the hypermethylation of its promoter.36, GPCR Compound Library 38 In the current study, the methylation level of the CDH1 promoter region, which we measured, was increased from 0% to 23.8% in HepG2 cells. From these results, we can see that the TSG methylation levels increased, regardless of the initial methylation status. The relative mRNA level measurement showed that GSTP1 expression was significantly decreased after transfection of the miR-152 inhibitor in comparison with the control group, whereas the CDH1 mRNA level was not click here significantly changed. This probably occurred because the increasing DNA methylation level of the CDH1 promoter was not sufficient to inhibit the mRNA expression. The hypermethylation of CpG islands of TSGs promotes oncogenesis not only through transcriptional inactivation of these genes but also through the following mechanisms: A signature CT mutation in cancer cells: the cytosine residues in the methylated dinucleotide CpG have a higher mutation rate than the unmethylated cytosine. The induction of chromosomal instability: aberrant DNA methylation leads to the genomic instability necessary for the development and progression of cancer, and DNA methylation

is also correlated with allelic deletions.41, 42 Moreover, HBV DNA has been shown to contain CpG islands that can be methylated in human tissue both in a nonintegrated form43 and after integration into the human genome.44 The methylation of viral CpG islands can regulate viral protein production,45 which likely reflects viral adaptation to host cells. A DNA methylation–associated blockade of viral antigen presentation could help the virus to evade our immune system. The depletion of DNMT1 and DNMT3B by siRNA or upon treatment with the DNA demethylation agent caused DNA hypomethylation of the HBV genome in HCC cells.46 In the present study, we have demonstrated that HBx can up-regulate DNMT1 activity by inhibition of miR-152. These mechanisms may also be involved in the methylation of the HBV genome and the survival of HBV in host cells.

In the absence of artificial organ

support, failure of the hepatic graft to promptly function would be tantamount to death. Finally, how could immediately life-supporting deceased donor livers be obtained in an era in which death was defined as the cessation of heartbeat and respiration? These questions and issues mandated consideration of the less draconian auxiliary hepatic transplant operation of Welch that might allow recipient survival, even if the graft failed. This option was undermined when the rapid atrophy of auxiliary livers that previously had been ascribed to rejection in unmodified dogs,86,113 was shown to be equally severe in animals in which rejection was prevented with azathioprine.11 The die was cast for the liver replacement (orthotopic) option. Liver replacement was carried out in seven deceased donor liver recipients between March 1963 and January 1964: BIBW2992 mw five in Denver (cases 1-4 and 6), one in Boston (case 5 by Moore’s team), and one in Paris (case 7) (Table 3).10,

1188,114 All seven patients died, two during the operation and the other five after 6.5-23 days. Neither primary nonfunction nor uncontrolled rejection of the grafts were lethal factors in any selleck antibody inhibitor of the failures. At autopsy of the four Denver patients who survived the operation, pulmonary emboli were found that apparently had originated in the bypass tubing used to decompress the blocked systemic and splanchnic venous beds during the removal and replacement of the native liver. Ironically, the bypass which had been an essential component of the canine operation, is not mandatory in most human recipients, or even in dogs if venous collateralization is

encouraged by bile duct ligation a month in advance.115 By the time our fourth and fifth liver recipients were find more reported to the American Surgical Association in April 1964,11 all clinical liver transplant activity had ceased in what would be a voluntary 3.5-year worldwide moratorium. The self-imposed decision to stop did little to quiet polite but unmistakably disapproving discussions of an operation that had come to be perceived as too difficult to ever be tried again. In effect, it now would be necessary to return to ground zero and reexamine all five of the themes of Table 1. The central assumption of Theme I had been that portal venous blood contained hepatotrophic molecules. The hypothesis was consistent with our results in 1958-1960 in nonimmunosuppressed canine recipients of replacement livers,3 and especially with the acute atrophy of Welch’s auxiliary grafts in azathioprine-treated dogs (see above, and Starzl et al.11). The possibility was now explored of providing the auxiliary allografts with direct access to the portal molecules.116 But what were the hepatotrophic factors? Using double liver fragment nontransplant models derived from Welch’s auxiliary liver operation (Fig.

In the absence of artificial organ

support, failure of the hepatic graft to promptly function would be tantamount to death. Finally, how could immediately life-supporting deceased donor livers be obtained in an era in which death was defined as the cessation of heartbeat and respiration? These questions and issues mandated consideration of the less draconian auxiliary hepatic transplant operation of Welch that might allow recipient survival, even if the graft failed. This option was undermined when the rapid atrophy of auxiliary livers that previously had been ascribed to rejection in unmodified dogs,86,113 was shown to be equally severe in animals in which rejection was prevented with azathioprine.11 The die was cast for the liver replacement (orthotopic) option. Liver replacement was carried out in seven deceased donor liver recipients between March 1963 and January 1964: Epigenetics inhibitor five in Denver (cases 1-4 and 6), one in Boston (case 5 by Moore’s team), and one in Paris (case 7) (Table 3).10,

1188,114 All seven patients died, two during the operation and the other five after 6.5-23 days. Neither primary nonfunction nor uncontrolled rejection of the grafts were lethal factors in any Metformin of the failures. At autopsy of the four Denver patients who survived the operation, pulmonary emboli were found that apparently had originated in the bypass tubing used to decompress the blocked systemic and splanchnic venous beds during the removal and replacement of the native liver. Ironically, the bypass which had been an essential component of the canine operation, is not mandatory in most human recipients, or even in dogs if venous collateralization is

encouraged by bile duct ligation a month in advance.115 By the time our fourth and fifth liver recipients were selleck reported to the American Surgical Association in April 1964,11 all clinical liver transplant activity had ceased in what would be a voluntary 3.5-year worldwide moratorium. The self-imposed decision to stop did little to quiet polite but unmistakably disapproving discussions of an operation that had come to be perceived as too difficult to ever be tried again. In effect, it now would be necessary to return to ground zero and reexamine all five of the themes of Table 1. The central assumption of Theme I had been that portal venous blood contained hepatotrophic molecules. The hypothesis was consistent with our results in 1958-1960 in nonimmunosuppressed canine recipients of replacement livers,3 and especially with the acute atrophy of Welch’s auxiliary grafts in azathioprine-treated dogs (see above, and Starzl et al.11). The possibility was now explored of providing the auxiliary allografts with direct access to the portal molecules.116 But what were the hepatotrophic factors? Using double liver fragment nontransplant models derived from Welch’s auxiliary liver operation (Fig.

This view is further supported by the observation that siRNA-mediated suppression of c-Src expression by 71 ±

4% lowered the half maximal inhibitory concentration (IC50) of herbimycin A to a similar extent from 0.11 μM to 0.038 μM (Fig. 2D). This inhibition of HCV replication upon suppression of c-Src expression by specific siRNA could be rescued by expression of neither Yes nor Fyn (Supporting Information Fig. 3). Thus, the two other ubiquitously expressed Src family members Yes and Fyn are not able to substitute c-Src. According to this, knockdown of Yes and Fyn by siRNA did not largely affect viral protein expression (Supporting Information Fig. selleck inhibitor 4). In summary, these data suggest that, from those Src family members that are ubiquitously expressed, c-Src plays a relevant role for HCV replication, whereas Fyn and Yes seem to be dispensable. Because herbimycin A and c-Src siRNAs significantly affected the abundance of viral genomic RNA, we raised the question of whether c-Src binds to the viral RNA-dependent RNA polymerase (NS5B). As shown in Fig. 3A, NS5B could be coprecipitated with c-Src–specific antibodies

from whole protein extracts prepared from Huh 9-13 cells harboring the subgenomic HCV replicon. Accordingly, in pull-down assays using GST-tagged c-Src, LY294002 in vivo NS5B could also be precipitated from cell lysates prepared from replicon-expressing Huh 9-13 cell lines (Fig. 3B) or from Huh cell lines infected with two different JFH1-derived viral HCV strains (Supporting Information Fig. 5). Conversely, GST-tagged NS5B was also able to precipitate c-Src (Fig. 4). Apart from confirming the assumption that NS5B interacts with c-Src, the pull-down assays using GST-tagged c-Src further indicated that NS5A

also binds to c-Src. These data suggest that either a protein complex this website comprising c-Src, NS5A, and NS5B is formed or two independent complexes comprising c-Src plus NS5A or c-Src plus NS5B (Figs. 3 and 4). To define the regions of c-Src that are required for the interaction with NS5A and NS5B in more detail, GST-tagged deletion mutants of c-Src were constructed and used for pull-down assays. As demonstrated in Fig. 3B, c-Src deletion mutants lacking the SH3 domain were unable to coprecipitate NS5B, whereas coprecipitation of NS5A was reduced but not abrogated. In contrast, deletion of the SH2 domain completely interrupted the interaction of c-Src with NS5A, but did not affect the interaction with NS5B. This indicates that the interaction of c-Src with NS5A requires the SH2 domain, whereas the interaction with NS5B depends on the presence of the SH3 domain. Pull-down assays using isolated GST-tagged SH3 domains of c-Src, Fyn, Hck, Lck, and c-Abl (Fig.