Figure 6. Other Smac-Mimetic-Sensitive Cells Respond in a Similar Fashion as HCC461 Cells.
Cell lines HCC44, MDA-MB-231, and SK MEL-5 were tested for rescue by siRNA transfection and neutralizing antibody pretreatments. siRNAs targeting caspase-8, TNFR1, TNFα, and RIPK1 (Dharmacon siGENOME SMARTpool predesigned pools of four oligos), as well as neutralizing antibody against TNFα (R&D Systems), were able to rescue these cell lines from Smac-mimetic-mediated apoptosis, while caspase-9 siRNA transfection (Dharmacon siGENOME SMARTpool) and pretreatment with TRAIL (Biolegend) neutralizing antibody had no rescue effects.
(A) HCC44.
(B) MDA-MB-231.
(C) SK MEL-5.
(D) H2009 cells were shown to respond to 100 nM compound 3 and 100 ng/ml TNFα. siRNA knockdown of caspase-8, TNFR1, and RIPK1 were able to rescue these cells from compound 3/TNFα cotreatment. siRNA transfection and cell viability assays were done as previously described. Each graphical representation indicates the mean ± SD of at least three independent testing conditions.