Fig. 4
From: Tumor suppressor SMAR1 regulates PKM alternative splicing by HDAC6-mediated deacetylation of PTBP1
SMAR1-mediated PKM alternative splicing is HDAC6 dependent. a Expression of PKM isoforms upon shRNA-mediated knockdown of HDAC6 in MCF7 by western blot. b Expression of PKM isoforms upon SMAR1 overexpression followed by Tubacin (5 μM) treatment in MCF7 by western blot. c Confocal microscopy to check the expression of eGFP and mCherry in MCF7 cells transfected with PKM minigene along with Flag-SMAR1. Relative fluorescence intensity of eGFP and mCherry was quantified with the help of ImageJ and fold change in eGFP/mCherry ratio was calculated for control, control + Tubacin (5 μM), Flag-SMAR1 and Flag-SMAR1 + Tubacin (5 μM). Fold change in eGFP/mCherry ratio for SMAR1 overexpression and Tubacin treatment validates the role of HDAC6 in SMAR1-mediated regulation of PKM alternative splicing (n = 3). Error bars show mean values ± SD. Differences were considered statistically significant with *p < 0.05, **p < 0.01, and ***p < 0.001; ns non-significant difference (p > 0.05)