Fig. 1
RSL1D1 Regulates the Proliferation and Survival of HCT116 Colorectal Cancer Cells. HCT116p53+/+ and HCT116p53−/− cells were transfected with siRSL1D1 to downregulate RSL1D1 expression. The cells transfected with siNC were used as a negative control. A The mRNA levels of RSL1D1 were determined by qRT-PCR analysis. GAPDH was used as an internal control to normalize the values. The normalized value of siNC-treated HCT116p53+/+ cells was set to 1. B The levels of RSL1D1 protein were determined by western blot analysis. β-actin was used as a loading control. C Cell proliferation was evaluated by MTT assay. The cells were seeded to a 96-well plate and incubated for 0, 1, 2, and 3 days. The values on day 0 were normalized to 1. D Cell cycle was determined by PI staining. The stained cells were subjected to flow cytometry to analyze the average percentage of each cell cycle phase. Sub-G1 indicates the apoptotic cell population. E Cell apoptosis was determined by Annexin V-FITC/PI double staining. The stained cells were subjected to flow cytometry to analyze the percentage of apoptotic cells. The cells in the right lower (Annexin V-FITC+/PI−) and right upper (Annexin V-FITC+/PI+) quadrants indicate early and late apoptosis, respectively. A, C-E Data are represented as mean ± SD. Student’s t test. *P < 0.05 and **P < 0.01 denote significant difference