Western Blot Analysis of MBD3 Knockdown

Western blots to confirm MBD3 knockdown were performed as previously described [22 (link)]. In brief, cells expressing shRNAs for 7 days were acquired by cell sorting, lysed and proteins resolved by SDS–PAGE. Blotting was performed by using a wet-tank blotting system (BioRad, Hercules, CA, USA), and membranes were stained with primary antibodies anti-MBD3 (1:5000), anti-histone H3 (1:2000) or anti-β-actin (1:5000). Membranes were stained with secondary antibodies IRDye^® 800CW (LI-COR Biosciences, Lincoln, NE, USA) or IRDye^® 680RD (LI-COR Biosciences, Lincoln, NE, USA) (1:15,000) and imaged by using the Odyssey Infrared Image System (LI-COR Biosciences, Lincoln, NE, USA).