, For protein extraction, cells were rinsed with ice-cold PBS and lysed for 30 min at 4 ? C in RIPA lysis and extraction buffer (Sigma) supplemented with a protease/phosphatase inhibitor cocktail (Sigma-Aldrich, St. Quentin Fallavier, France), 5 mM sodium orthovanadate, For IGF-1 stimulation, the cells were harvested overnight before IGF-1 addition into an FBS-free culture medium, pp.5-21078
, After washing, blots were incubated for 1 h with a horseradish peroxidase-linked anti-rabbit antibody (Sigma-Aldrich, St. Quentin Fallavier, France) and processed for a chemiluminescent substrate, Antibodies for detection of P-IGF1R? (Tyr1131) (#3021), p.2
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