Research Paper Volume 12, Issue 16 pp 16083—16098

Figure 3. Chidamide has cytostatic and cytotoxic effects on CLL cells. (A) Flow cytometry using Annexin V–FITC/PI staining for cell apoptosis analysis. Primary CLL cells were incubated with 4μmol/L chidamide for 24 hours. Representative data shown are from 9 patients and three independent experiments of MEC-1, JVM-3 respectively. The bar graphs showed the percentage of apoptotic cells. (B) Immunoblotting analysis of poly (ADP-ribose) polymerase (PARP) in primary CLL cells after chidamide (CHI, 4μmol/L) treatment for 24 hours. Shown are 3 representative blots from the samples of 6 patients. The bar graph represents the relative PARP cleavage/GAPDH ratio measured by immunoblotting. (C) CCK8 assay for detecting metabolically active cells. Primary CLL cells were incubated with different concentrations of chidamide for 24 and 48 hours. Viability of cells compared with the corresponding controls was shown from three independent experiments respectively. (D) Cell apoptosis assay as described in (A). MEC-1, JVM-3 cell lines were incubated with indicated concentrations of chidamide for 24 and 48 hours. (E, G) Immunoblotting analysis of PARP as described in (B). MEC-1 and JVM-3 cell lines were incubated with indicated concentrations of chidamide for 24 hours. (F, H) CCK8 assay as described in (C). MEC-1, JVM-3 cell lines were incubated with different concentrations of chidamide for 24 and 48 hours.