As previously described in 33 (link), 50 μg of total protein was electrophoretically separated using a 4–20% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane (Bio-Rad, Hercules, CA, USA). Membranes were blocked in Odyssey blocking buffer (LI-COR, Lincoln, NE, USA) for 1 hr at room temperature. Membranes were probed with primary antibodies overnight at 4°C: rabbit anti-MeCP2 (1:1000, Millipore cat no. 07–013, Burlington, MA, USA) and mouse anti-Gapdh (1:1000, ThermoFisher cat. no. MA5–15738, Waltham, MA, USA), followed by the fluorescent secondary antibodies: goat anti-rabbit (800nm, 1:5000, LI-COR, Lincoln, NE, USA) and goat anti-mouse (680nm, 1:10,000, LI-COR, Lincoln, NE, USA). Fluorescence was detected using the Odyssey (LI-COR, Lincoln, NE, USA) imaging system at the Vanderbilt University Medical Center Molecular Cell Biology Resource (MCBR) Core and then quantified using the Image Studio Lite software (LI-COR, Lincoln, NE, USA). Values were normalized to Gapdh and compared relative to wild-type controls.