Figure 2 | Cell Death & Disease

Figure 2

From: Neoalbaconol induces energy depletion and multiple cell death in cancer cells by targeting PDK1-PI3-K/Akt signaling pathway

Figure 2

NA triggers necroptosis in cancer cells. (a) C666-1 cells were treated with NA (40 μM), paclitaxel (200 nM) or DMSO. The effects on morphology of C666-1 cells were detected by optical microscopy. All panels are of the same magnification ( × 100 or × 40) and each panel is representative of three experiments. (b) Nec-1(50 μM) was preincubated with NA-treated cells. Cell viability was analyzed by the MTS assay. Data shown are the mean+S.D. of three experiments. *P<0.05. **P<0.001. (c) Immunoblotting analysis was adopted to investigate the effect of NA on the expression level of RIP1 and RIP3. β-Actin served as a loading control. (d) A Flag-tagged RIP3 plasmid was transfected into C666-1 cells. A coimmuoprecipitation assay was used to analyze the effect of NA (40 μM), Nec-1(50 μM) or their combination on the interaction of RIP1 and RIP3. (e) The interaction of endogenous RIP1 and RIP3 was analyzed by an immunofluorescence confocal assay. Statistical analyses of the percentage of cells contain four fields. All panels are of the same magnification ( × 1600) and each panel is representative of three experiments. Data shown are the mean+S.D. *P<0.05. **P<0.001

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