Fig 1: Calcyclin-binding protein (CACYBP) knockdown facilitated degradation of minichromosome maintenance complex component 2 (MCM2) by inducing ubiquitination. (A) Relative mRNA levels of the top eleven differentially expressed genes (DEGs) in HUCCT1 cells after CACYBP silencing as determined by qPCR analysis. (B) Relative protein levels of MCM2, NR1H3, PRIM1, CCND1 and CCNG1 in HUCCT1 cells after CACYBP silencing as determined by western blot assays. (C) Viability of HUCCT1 cells after knockdown of NR1H3, PRIM1, CCND1, CCNG1 and MCM2 as assessed by Celigo cell count assays at indicated times. (D) Relative protein level of MCM2 in CACYBP-knockdown HUCCT1 cells treated with cycloheximide (50 mg/mL). (E) Relative protein level of MCM2 in CACYBP-knockdown HUCCT1 cells exposed to 10 µmol/L proteasome inhibitor MG-132 treatment. (F) Ubiquitination of MCM2 immunoprecipitated by anti-MCM2 antibody. Data are represented by mean ± standard deviation. (G) Detection of K48 and K63 mediated ubiquitination of MCM2 in HUCCT1 cells. *p < 0.05, **p < 0.01, ***p < 0.001.