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. 2021 Dec 6;75(6):1386–1401. doi: 10.1002/hep.32177

FIGURE 4.

FIGURE 4

HIF‐1α and SREBP1 mediating increased glucose to CL anabolism represses cytochrome c release in radioresistant HCC cells. (A) Restorative effect of M‐HCl (20 μM, 48 h) on IR‐induced apoptosis and cytochrome c release in the indicated cell lines. Glyceraldehyde 3‐phosphate dehydrogenase and translocase of outer mitochondrial membrane 20 were loaded as cytoplasmic and mitochondrial markers, respectively. (B) Relative CL content (left) and cell survival determined by MTT assays (right) of radioresistant cells transiently expressing either scrambled siRNA or siRNA against CRLS1 following IR treatment. (C) Western blot of downstream targets in IR‐R cells upon HIF1A or SREBP1 knockdown using shRNA. (D) Cell viability determined by MTT assays in resistant cells upon HIF1A or SREBP1 knockdown. (E,F) Restorative effect of M‐HCl (E) or CRLS1 overexpression (F) on IR‐induced apoptosis in radioresistant cell lines with HIF1A or SREBP1 knockdown. Survival data were normalized to those of unirradiated control cells. Data are represented as mean ± SEM of at least three replicates. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Abbreviations: Cyto, cytoplasmic; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; DAPI, 4‐6‐diamidino‐2‐phenylindole; (M), mature; Mito, mitochondrial; ns, not significant; (P), precursor; TOM20, translocase of outer mitochondrial membrane 20 [Color figure can be viewed at wileyonlinelibrary.com]