HDAC Inhibitor Modulates DNMT1 in Melanoma

Two melanoma cell lines (MH-0331 and WP-0614) were treated for 24 h with HDAC inhibitor panobinostat (Cat# HY-10224, MedChemExpress, Monmouth Junction, NJ, USA) at different concentrations (10, 50, and 100 nM). Protein was then extracted and evaluated by Western blot for DNMT1, ac-DNMT1, and β-actin as previously described [38 (link),41 (link)], except for the Abs utilized, which were the polyclonal rabbit anti-human DNMT1 Ab (1:100 dilution, Cat# NB100-264, Novus Biologicals), the rabbit anti-human ac-DNMT1 Ab (1:500 dilution, Cat# A5595, ABclonal), and the mouse monoclonal anti-human β-actin Ab (1:10000 dilution, Cat# A5441, Sigma-Aldrich). All Western blot images were analyzed with ImageJ software (http://imagej.nih.gov/ij/ (accessed on 3 August 2021). All the uncropped Western blot images were included in Figure S1.