Fig. 8: SH003-induced autophagy is regulated via G9a. | Cell Death & Disease

Fig. 8: SH003-induced autophagy is regulated via G9a.

From: SH003 activates autophagic cell death by activating ATF4 and inhibiting G9a under hypoxia in gastric cancer cells

Fig. 8

a Protein levels of H3K4me2, H3K9me2, H3K9me3, H3K27me2, and H3K27me3 were investigated by Western blotting in nuclear fraction samples isolated from SH003 (0, 200, and 400 μg/mL, 24 h)-treated AGS and SNU-638 cells, and H3 was used as a protein loading control. Protein levels of G9a and p-STAT3 were detected by Western blot assay in SH003-treated AGS and SNU-638 cells and β-actin was used as a protein loading control. b, c The localization of the G9a and ATF4 on the LC3B promoter. SH003 regulates G9a and ATF4 binding at the LC3B promoter. SH003 (400 μg/Ml, 24 h) treatment was performed in AGS and SNU-638 cells, and real-time ChIP assays of the LC3B promoter region (+541 to +656) were performed with G9a and ATF4 antibody. d, e AGS and SNU-638 cells were transfected with control or G9a siRNA in the presence or absence of SH003 (400 μg/mL, 24 h), and then, cell viability tests, LDH assay, and Western blotting were performed; *p < 0.05. f, g AGS and SNU-638 cells were pretreated with 3-MA (5 mM) for 4 h, and then, were treated with SH003 (400 μg/mL, 24 h) and/or BIX-01294(10 μM, 24 h). Cell viability, LDH release, and Western blot analyses of G9a and LC3B in the AGS and SNU-638 cells treated with SH003 + BIX-01294 + 3-MA were performed. β-actin was used as a protein loading control; *p < 0.05.

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