As previously described in Reference 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 h 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 (800 nm, 1:5000, LI‐COR, Lincoln, NE, USA) and goat anti‐mouse (680 nm, 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.