Figure 6.

MIR145-3p mimic enhances the anti-MM activity of bortezomib through autophagy activation. (A) LP-1 cells were transfected with MIR145-3p or MIR control (MIRctrl) for 72 h, and then treated with 20 nM bortezomib (Bort) or vehicle for 24 h. Cell death was measured by ANXA5-7-AAD staining. The percentage of ANXA5-positive cells was presented as mean ± SD from 3 independent experiments. (B) LP-1 cells were transfected with siRNA-HDAC4 or negative siRNA control (sictrl) for 72 h, and then treated with 20 nM Bort or vehicle for 24 h. Cell death was measured by ANXA5-7-AAD staining. The percentage of ANXA5-positive cells was presented as mean ± SD from 3 independent experiments (all *P < 0.05; **P < 0.01). (C) LP-1 cells were transfected with MIR145-3p mimic, MIRctrl, siRNA-HDAC4 or sictrl for 72 h, and then treated with 20 nM Bort or vehicle for 24 h. MAP1LC3B and SQSTM1 protein levels were determined by western blotting. GAPDH was used as loading control. The experiments were performed in triplicate. (D) LP-1 cells were transfected with MIR145-3p mimic or MIRctrl for 72 h, in the presence or absence of 20 nM Bort, followed with or without BAF (20 nM) treatment for 4 h. MAP1LC3B and SQSTM1 protein levels were determined by western blotting. GAPDH was used as loading control. The experiments were performed in triplicate.