Research Paper Volume 12, Issue 16 pp 16083—16098

Figure 2. Chidamide inhibits autophagy in mTOR-independent way by post-transcriptionally regulation in CLL cells. (A) The effect of chidamide on the PI3K/AKT/mTOR pathway was assessed in MEC-1 cells by immunoblotting through the analysis of the phosphorylation status of mTOR during time course in presence or absence of chidamide (CHI, 16μmol/L). The bar graph showed the relative expression level of p-mTOR/mTOR with respect to the control groups. (B) The effect of chidamide on modulating autophagy gene expression in transcriptional level. MEC-1 cells were treated with chidamide (CHI, 16μmol/L) for 8 or 18 hours, then autophagy-related genes LC3, SQSTM1, ATG7, and ATG3 were determined by qRT-PCR. ΔCt was used to measure statistical significance and the results were shown in the bar graphs. (C, D) MEC-1 cells were treated with chidamide (CHI, 8 or 16μmol/L) alone or in presence of MG132 (20μmol/L) or cycloheximide (CHX, 20μmol/L), collected after 24 hours and immunoblotted with antibodies against LC3 and SQSTM1. The bar graphs showed the expression level of proteins with respect to the control groups.