Fig. 7
Effects of corosolic acid on various phenotypes of hepatocellular carcinoma cells (A) Hepatocellular carcinoma cells Bel-7404 and HepG2 were treated with different concentrations of corosolic acid (2, 4, 8, 16, 32, 64, 128µM) for 24 h, and CCK-8 assay was performed to detect the cytotoxicity of candidate drugs. IC50 values were calculated to evaluate the cytotoxicity of candidate drugs. (B) SYTOX-Green staining was used to detected Cell death after treatment with different concentrations of corosolic acid (0, 10, 20, 30µM) for 24 h. (C) Colony formation in Bel-7404 and HepG2 cells after 7 days, the colonies were treated with corosolic acid (0, 15, 30µM) for 24 h and then were fixed with 4% paraformaldehyde as well as stained with 0.1% crystal violet. (D) PI staining was used to detect the effect of corosolic acid (0, 10, 20, 30µM) for 24 h. Flow cytometry was used to detect the cell cycle distribution of Bel-7404 and HepG2 cells (E) Western blot was used to determine the expression level of P4HA2 protein in Bel-7404 and HepG2 cell lines after corosolic acid (0, 15, 30µM) treatment for 24 h. In order to save and recycle antibodies and save time, membranes were cut in horizontal strips at molecular weight ranges for target proteins and the information was shown in the Supplementary File. Data are shown as mean ± SD of three biological replicates. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.001 indicates statistical significance