Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

Figure 3 Impact of suberoylanilide hydroxamic acid treatment on the level of endoplasmic reticulum-phagy and autophagy-mediated cell death in Huh7 and MHCC97L cells. A-C: Huh7 and MHCC97L cells were exposed to 0, 1, 3, 6, and 12 μmol/L suberoylanilide hydroxamic acid for 48 h. Proteins related to the reticulophagy pathway were detected using western blotting. Relative protein expression levels were normalized to β-actin; D: The colocalization coverage of the endoplasmic reticulum and lysosomes were observed using a confocal microscope (× 400). Representative pictures from three independent replicate assays are exhibited; E: Huh7 and MHCC97L cells were pretreated with 0.5 nmol/L BafA1 for 12 h, followed by 12 μmol/L suberoylanilide hydroxamic acid treatment for 48 h. Hoechst 33342/propidium iodide (PI) double staining localization assay was utilized to observe nuclear coagulation. After Hoechst 33342/PI double staining, the death rate of hepatocellular carcinoma cells was quantified based on the ratio of pink cells/blue cells. Experiments were repeated thrice. Representative pictures from three independent replicate assays are shown. Data are exhibited as mean ± standard deviation. ^aP < 0.05 vs 0 μmol/L group. SAHA: Suberoylanilide hydroxamic acid; ER: Endoplasmic reticulum; FAM134B: Family with sequence similarity 134 member B.