Figure 3
C646 inhibits expressions of G2/M cell cycle-associated genes. (A, B) Expression profiles of molecules associated with the G2/M transition at the protein level (A) and mRNA level (B) following C646 treatment. mRNA expression of cyclin B1 and CDK1 was inhibited after 48-h treatment with 30 µM C646 treatment in PSN1 and MIAPaCa2 cells. Inhibition of protein expression of cyclin B1 and CDK1 after 72-h C646 treatment was confirmed by Western blotting. Phosphorylated histone H3 (Ser10), recognized as an M phase marker, was also inhibited by C646 treatment. Error bars represent mean ± SD. *p < 0.05 vs DMSO vehicle control. (C) Dose-dependent analysis of G2/M cell cycle-associated molecules following C646 treatment (10–50 µM) by Western blotting in PSN1 and MIAPaCa2 cells. (D) Time-dependent analysis of histone H3 acetylation and G2/M cell cycle-associated molecules following C646 treatment for 24 and 48 h in PSN1 cells. (E) Quantitative ChIP analysis of cyclin B1 and CDK1 with C646 treatment (40 µM) in PSN1 cells. H3K9ac and H3K27ac levels at the promoter region of cyclin B1 and CDK1 were significantly decreased by C646 treatment. Error bars represent mean ± SD. *p < 0.05 vs controls.
