Western blot of nuclear hormone receptors in A375(DRO) resistant cell lines. 3a: 60 μg of nuclear protein extract from the resistant A375(DRO) sublines was size-separated on a 10% SDS-PAGE gel and transferred to nitrocellulose. The blot was blocked with 10% nonfat milk and incubated with RXRγ (MS-1343-P NeoMarkers) and RXRα (sc D-20) antibodies at a concentration of 1:500 and PPARγ (H-100) rabbit polyclonal ab (sc-7196, Santa Cruz Biotechnology, Santa Cruz, CA) at 1:500. Secondary antibodies were anti-rabbit IgG conjugated to horse-radish peroxidase at a 1:5000 dilution for RXRs and 1:1000 for PPARγ (GE Healthcare UK). β-Actin was measured as a loading control. 3b: PPARγ receptor to β-Actin ratio was calculated using an Alpha Innotech alpha imager.