FIG. 5. .

Effects of caveolae disruption with MβCD on FGF2-induced FRS2α-GRB2-GAB1 formation in oFPAE cells. A) The cells were pretreated with or without 10 mM MβCD for 1 h, followed by stimulation with 10 ng/ml FGF2 for 5 min. Cells were lysed to isolate the caveolae membranes by sucrose-gradient fractionation. The caveolae membrane fractions were analyzed by immunoblotting with antibodies for FRS2α, GRB2, GAB1, and cav-1. B) The cells were pretreated with or without 10 mM MβCD for 1 h, followed by stimulation with 10 ng/ml FGF2 for 5 min. Total cellular proteins were prepared for IP with anti-GAB1 antibody, followed by immunoblotting with antibodies for FRS2α, GRB2, and GAB1. Experiments were repeated four times using cells from different pregnant ewes, and band intensities were quantified and expressed as mean ± SD. Bars with different superscripts differ significantly (P < 0.05).