Protein Isolation and Western Blot Analysis

Protein (either nuclear and cytoplasmic fractions or total extracts) was isolated using the Nuclear Extract Kit (Active Motif, La Hulpe Belgium) according to the manufacturer’s protocol. The protein concentration was measured using the Bio-Rad Protein Assay (Bio-Rad, Hercules, California, USA) according to the manufacturer’s protocol. Western blot was performed using the anti-GAPDH rabbit mAb (14C10; Cell Signaling Technology, Leiden, The Netherlands), the anti-α-tubulin mAb (DM1A; Calbiochem, EMD Chemicals Inc San Diego, USA), the anti-histone H3 (ab1791; Abcam, Cambridge UK), the anti-SNAIL mouse mAb (L70G2; Cell Signaling), the anti-MYOD rabbit pAb (M-318; Santa Cruz, CA, USA), the anti-HDAC1 rabbit pAb (Poly6074 clone; Biolegend, San Diego CA, USA), the anti-HDAC2 mouse mAb (Biolegend, 3F3/HDAC2), anti-myogenin mouse mAb (F5D, sc-12732, Santa Cruz Biotechnology), anti-fast myosin skeletal heavy chain mouse mAb antibody (MyHC, MY-32, ab51263, Abcam), and secondary anti-rabbit and anti-mouse antibodies conjugated with horseradish peroxidase (HRP, Santa Cruz Biotechnology) as previously described^{24 (link)}.