DR4 and DR5, also known as TRAIL R1 and chemical library , respectively, incorporate useful cytoplasmic Wnt Pathway death domain motifs, which keep company with Fas associated death domain protein upon activation by apoptotic signs such as TRAIL. FADD contains the death effector domain and is involved in the activation of caspase 8. Therefore, increased surface expression of DR4 and DR5 observed in I3M treated cells might subscribe to the caspase 8 activation observed in Fig. 2A. It has been reported that expression of DR4 or DR5 is transcriptionally regulated by p53 tumor suppressor gene. In this study, the significantly elevated p53 and p21 protein level in I3M treated cells suggests the possibility that I3M encourages DR4 and DR5 expression via activation of p53. It has also been reported that in HeLa cells p53 could be functionally upregulated as evidenced by the increase of p21 protein, even though lots of previous studies have shown that HeLa cells are either p53 deficient or with minimal expression level of p53. In reality, therapy using other indirubin types have been observed to up manage p53 in human cancer cells, implying a standard procedure in indirubin derivativeinduced apoptosis. At present, it remains to be further examined as how I3M causes p53 accumulation and activation. Yet another possible mechanism by which death receptor is promoted by I3M mediated apoptosis is through modulation of NF kB task. The anti apoptotic functionality of NF kB has been well established via the transcriptional regulation of numerous anti apoptotic genes such as for instance. Indirubin and its derivatives have now been reported to prevent the NF kB signaling pathway stimulated by various activators, including TNFa, PMA and H2O2. In this study, I3M didn’t affect the basal level of NF kB transcriptional activity. It remains to be Plastid further studied whether I3M mediated caspase 8 activation is achieved via the reduction of the NF kB signaling pathway. On another hand, I3M induced apoptosis in HeLa cells also display a reply typical of type II cells, since the intrinsic mitochondrial route as shown by caspase9 activation and cytochrome c release is mediated by Bid common compound library cleavage downstream of caspase 8 activation. More over, Bax conformational change does occur whilst the effects of caspase 8 activation and Bid cleavage centered on immunofluorescence and immunoprecipitation knowledge using conformation specific antibody 6A7. Along with BH3 only proteins, the anti apoptotic Bcl 2 family members will also be proven to regulate the pro apoptotic activity of Bax through sequestrating Bax by the forming of heterodimers. In the present study, ectopic expression of Bcl 2 protein provided moderate safety against I3M induced cell death.