Western Blot

Tissues were mechanically homogenized using a pellet pestle motor in lysis buffer (20 mM HEPES (pH 7.8), 100 mM NaCl, 5% glycerol, 0.5% NP-40, 5 mM EDTA, 1 mM dithiothreitol, and phosphatase and protease inhibitors), lysed by ultrasound sonication (Diagenode Bioruptor) and centrifuged for 10 min at 4°C. Total protein content was determined by Bradford quantification. Western blot analysis was performed as previously described⁷⁶. GAPDH or αTUBULIN served as loading controls. Images were acquired with Image Quant LAS 4000 (GE healthcare). The list of antibodies used in this study and the corresponding dilutions are provided in Supplementary Table 4.