Whole-cell lysate was harvested in boiling lysis buffer (25 mM Tris·HCl pH 7.6, 1% SDS, 1 mM EDTA, 1 mM EGTA) and 50 µg of total protein extract was loaded into an 8% acrylamide gel. The antibodies used were designed against ESRP1(Sigma-Aldrich, Milan, Italy) [55 (link)], ESRP2 (abcam ab113486), GAPDH (Santa Cruz Biotechnology, sc-32233, Dallas, TX, USA), and HSP90 (abcam ab59459).
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