Chromatin accessibility module identified by single-cell sequencing underlies the diagnosis and prognosis of hepatocellular carcinoma

Figure 6 DGRT1 regulated by chromatin accessibility promoted cell proliferation. A: Western blot analysis of DGAT1 and histone 3 lysine 27 acetylation expression in SNU449 cells treated with 1 μmol/L A485 (CREB-binding protein/P300 inhibitor) for 48 hours. Glyceraldehyde-3-phosphate dehydrogenase served as a loading control; B: Quantification of DGAT1 and H3K27Ac protein levels normalized to GAPDH (n = 3); C: Western blot of DGAT1 and proliferation marker in SNU449 cells transfected with DGAT1-targeting small interfering RNA. Small interfering RNA with non-targeting gene was used as control; D: Quantification of DGAT1 and proliferation marker levels normalized to GAPDH (n = 3); E: Representative images of 5-ethynyl-2’-deoxyuridine assay showing proliferating cells (EdU-positive, red) in DGAT1-knockdown SNU449 cells. Nuclei were stained with 4’, 6-diamidino-2-phenylindole, dihydrochloride (blue); F: Quantitative analysis of EdU-positive cells (n = 3), P value < 0.05; G: Cell counting kit-8 assay demonstrating reduced cell viability in DGAT1-knockdown SNU449 cells over 96 hours (n = 3), P value < 0.05. EdU: 5-ethynyl-2’-deoxyuridine; DAPI: 4’, 6-diamidino-2-phenylindole, dihydrochloride; DGAT1: Diacylglycerol acyltransferase-1; siDGAT1: Small interfering RNA targeting DGAT1; H3K27Ac: Histone 3 lysine 27 acetylation; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; PCNA: Proliferating cell nuclear antigen.