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The following day, membranes were washed with fresh 1 PBS with 0.1% Igepal and incubated for 45 min with gaot anti-mouse (ab6789, Abcam, 1:3,000) conjugated to horseradish peroxidase at a 1:3,000 dilution, and analyzed by enhanced chemiluminescence (ECL, Cytiva) using a Vilbert-Lourmat Photodocumentation Chemistart 5000 imager