Fig. 4

CREB1 promoted CCNA2 expression and enhanced HCC cell cycle progression depending on HERH-1. CREB1 was overexpressed in HCC cells with or without HERH-1 inhibition, and phenotypes of the HCC cells were detected to confirm the dependence of CREB1-assisted CCNA2 expression and function on HERH-1. a, b The endogenous CCNA2 level was measured by qRT-PCR (a, n = 2) and Western blot (b) assays. Full-length blots are presented in Fig. S8b. c, d The HCC cell viability and proliferation rate were detected by CCK-8 (c, n = 3) and doubling time (d, n = 3) assays. e, f DNA replication in HCC cells was detected by EdU staining and flow cytometry (e, n = 2) or fluorescence microscopy (f, n = 2). Scale bar, 20 μm. g The HCC cell cycle distribution was detected by PI staining and flow cytometry analysis (n = 2). *P < 0.05; **P < 0.01; n.s., not significant