Fig. 4.

Inhibition of HDAC activity releases miR-145-5p expression resulting in miR-145-5p target genes modulation. a RT-qPCR to evaluate the expression levels of miR-145-5p after 72 h of 3 mM VPA treatment of TC1889 cells. b Schematic representation miR-145-5p upstream genomic region (upper panel) and Chromatin Immunoprecipitation (ChIP) analysis (lower panel) to evaluate the histone H4 acetylation (H4Ac) status of R1 and R2 miR-145 upstream genomic regions (R1 located at -1500 bp and R2 located at -1000pb) after 24 h of 3 mM VPA treatment in TC1886 cells. The intronic region of CCNB1 gene (Int B1) was used as negative control. c RT-qPCR to evaluate the expression levels of Golm-1, EGFR, CDH2 and Psat-1 after 72 h of 3 mM VPA treatment of TC1889 cells. d Wester blot analysis to evaluate Golm-1, EGFR and SYP after 24 h and 48 h of 3 mM VPA treatment of TC1889 cells. e Morphological and IHC analyses for Golm-1, EGFR, CDH2 and Psat-1 after 72 h of 3 mM VPA treatment of TC1889 cells. P-value was calculated by unpaired t-test and a value of P ≤ 0.05(*), P ≤ 0.01(**) and P ≤ 0.001(***) was considered statistically significant