Fig. 3: PUMA is required for BRD9 inhibition-induced apoptosis in GIST. | Cell Death & Disease

Fig. 3: PUMA is required for BRD9 inhibition-induced apoptosis in GIST.

From: RETRACTED ARTICLE: BRD9 inhibition promotes PUMA-dependent apoptosis and augments the effect of imatinib in gastrointestinal stromal tumors

Fig. 3

A GIST-882 cells were treated with GSK602 at indicated concentrations for 24 h. Left, PUMA expression was analyzed by western blotting. Right, PUMA mRNA level was analyzed by real-time PCR. B GIST-T1 cells were treated with GSK602 at indicated concentrations for 24 h. Left, PUMA expression was analyzed by western blotting. Right, PUMA mRNA level was analyzed by real-time PCR. C GIST-882 cells were treated with GSK602 at indicated concentrations for 24 h. The expression of indicated Bcl-2 family members was analyzed by western blotting. D GIST-T1 cells were treated with GSK602 at indicated concentrations for 24 h. The expression of PUMA was analyzed by western blotting. E WT and PUMA-KO GIST-882 cells were treated with 0.5 μM GSK602 for 24 h. Apoptosis was analyzed by fragment nuclei assay. F WT and PUMA-KO GIST-T1 cells were treated with 0.5 μM GSK602 for 24 h. Apoptosis was analyzed by fragment nuclei assay. G WT and PUMA-KO GIST-882 or GIST-T1 cells were treated with 0.5 μM GSK602 for 24 h. Indicated protein level was analyzed by western blotting. H Cytosolic fractions isolated from WT and PUMA-KO GIST-882 cells treated with 0.5 μM GSK602 for 24 hr were probed for cytochrome c by western blotting. β-actin and cytochrome oxidase subunit IV (Cox IV), which are expressed in cytoplasm and mitochondria, respectively, were analyzed as the control for loading and fractionation. I WT and PUMA-KO GIST-882 or GIST-T1 cell lines were treated 0.5 μM GSK602 for 24 h. Cell viability was analyzed by colony formation assay. Results were expressed as means ± SD of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.

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