VPA regulated miRNA expression through increased histone acetylation. (A) VPA treatment increased the global protein acetylation of LX2 cells as detected by western blot using an antibody against pan acetyl-lysine, the band for histone was about 15KD. (B) The transfection condition for LX2 cells was optimized by Cy3 labeled siRNA transfection control (Cy3-siTC), and a minimal concentration of 50nM was used in the present study according to transfection efficiency, ×200. (C) The expression of HDAC2 or 3 mRNA in siHDAC2 or 3 transfected LX2 cells was detected by qRT-PCR. (D) Knockdown of HDAC2 and HDAC3 by siRNAs enhanced the acetylation of Histone H3, detected by western blot using an antibody against acetylated lysine (K) 27 of histone H3 (AcH3K27). (E, F) The expression of VPA-miRNAs (E) and mRNAs of VPA-protein-encoding genes (F) in siHDAC2 and 3 co-transfected LX2 cells was detected by qRT-PCR, all expressions were compared to untreated control or non-targeting siRNA negative control (siNC), *** P < 0.001, ** P< 0.01, *P < 0.05.