Fig. 2: SNG-induced loss of viability in CRC cells is not associated with activation of classical cell death pathways. | Cell Death Discovery

Fig. 2: SNG-induced loss of viability in CRC cells is not associated with activation of classical cell death pathways.

From: Targeting oxeiptosis-mediated tumor suppression: a novel approach to treat colorectal cancers by sanguinarine

Fig. 2

A Cells were treated with the indicated concentrations of SNG for 16 h. Apoptosis was determined by Annexin V-FITC apoptosis detection kit. Cells were treated with B indicated concentrations of SNG (HT-29 for 16 h and CaCo-2 cells for 6 h) C HT-29 and CaCo-2 cells were treated with SNG for the indicated time period. Western blot analysis was performed. The signal intensities of western blot bands were normalized to actin of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, *p < 0.05, **p < 0.01, ***p < 0.001 and ns = no significance. The Blots shown here are representative of three independent experiments. D Cells were pretreated with z-VAD for 1 h followed by SNG treatment (HT-29 for 16 h and CaCo-2 for 6 h) and cell viability was measured by using MTT assay. Similar experiment in PC3 cells treated with SNG (3 μM) in the presence or absence of z-VAD were used as a positive control for apoptosis. Data shown are means ± SD (n = 3) (***p < 0.001 and ns = no significance). HT-29 cells were pretreated with z-VAD for 1 h followed by SNG treatment for a further 16 h, E Western blot analysis was performed. The signal intensities of western blot bands were normalized to actin of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, *p < 0.05, ***p < 0.001. The blots shown here are representative of three independent experiments and F apoptosis was determined by Annexin V-FITC apoptosis detection kit. G Cells were pretreated with GSK963, GSK872, and NSA for 1 h, followed by incubation with SNG (HT-29 for 16 h and CaCo-2 for 6 h). Cell viability was measured by using MTT assay. Similar experiment in HT-29 cells treated with TCZ in the presence or absence of NSA was used as a positive control for necroptosis. Data shown are means ± SD (n = 3) (**p < 0.01, ***p < 0.001 and ns = no significance). H Cells were treated with SNG for the indicated time period, and Western blot analysis was performed. The signal intensities of western blot bands were normalized to actin of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, *p < 0.05, **p < 0.01, ***p < 0.001 and ns = no significance. I HT-29 and CaCo-2 cells were pretreated with Baf A1 followed by incubation with SNG (HT-29 for 16 h and CaCo-2 for 6 h). Cell viability was measured by using MTT assay. Similar experiment in U87MG cells treated with SNG (3 μM) in the presence or absence of Baf A1 was used as a positive control for autophagy. Data shown are means ± SD (n = 3) (***p < 0.001 and ns = no significance). J HT-29 cells were pretreated with Baf A1 followed by incubation with SNG, and Western blot analysis was performed. The signal intensities of western blot bands were normalized to actin of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, **p < 0.01.

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