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Figure 2. RCC1 knockdown inhibits proliferation and induces G0/G1 phase arrest in CRC cells. (A) Basal RCC1 expression in nine CRC cell lines. Expression in HCT116 was normalized to 1, and relative levels in the other cell lines are shown at right (one-way ANOVA); (B) Top: Western blot analysis of RCC1 protein levels following knockdown in HCT116 cells and DLD-1 cells. Bottom: Quantification showing decreased RCC1 levels in knockdown groups (one-way ANOVA, P < 0.001, P < 0.05); (C) qRT-PCR analysis of mRNA levels in HCT116 and DLD-1 cells following knockdown (one-way ANOVA, P < 0.001); (D and E) CCK-8/MTT proliferation assays and colony formation assays showing reduced growth following RCC1 knockdown (two-way ANOVA, P < 0.001); (F and G) Flow cytometry-based cell cycle analysis of synchronized HCT116 cells (two-way ANOVA, P < 0.001). ns, not significant, ^***P < 0.001, ^*P < 0.05. scr was defined as no knockdown of RCC1 and used as a control. RCC1: Regulator of chromosome condensation 1; CRC: colorectal cancer; ANOVA: analysis of variance; qRT-PCR: quantitative reverse transcription-polymerase chain reaction.