fig3

Figure 3. RCC1 knockdown impairs proliferation in 5-FU- and Doxo-resistant HCT116 cells. (A and B) Validation of RCC1 knockdown in 5-FU- and Doxo-resistant HCT116 cells by western blotting (A) and qRT-PCR (B) (two-tailed unpaired t-test, P < 0.001); (C and D) Reduced proliferation (CCK-8 assay) and colony formation following RCC1 knockdown (two-way ANOVA, P < 0.001, P < 0.05); (E) Representative images of 5-FU- or Doxo-resistant CRC spheroids cultured for 5 days. Scale bar: 400 µm; (F) Total spheroid counts and number of spheroids with diameters > 75 μm (two-tailed unpaired t-test, P < 0.05); (G) Representative images of 5-FU- or Doxo-resistant CRC single spheroids cultured for 8 days. Scale bar: 400 μm; (H) Quantitative analysis of total single spheroids (two-tailed unpaired t-test, P < 0.05). ns, not significant; ^***P < 0.001; ^*P < 0.05. RCC1: Regulator of chromosome condensation 1; 5-FU: 5-fluorouracil; Doxo: doxorubicin; qRT-PCR: quantitative reverse transcription-polymerase chain reaction; ANOVA: analysis of variance; CRC: colorectal cancer.