Fig. 2. Effects of Src^527F/c-Yes chimeras on cellular phosphotyrosine. cell morphology, and the actin cytoskeleton. (A) 30 µg of day 12 RIPA lysates from mock-transfected CEF or cells expressing Src^527F, LA29, or the chimeric constructs were separated by 8% SDS-PAGE. Lysates were transferred to PVDF membrane, and probed with a rabbit anti-phosphotyrosine antibody Molecular weight markers are shown on the left side of the figure. Protein bands of note are highlighted by their M_r, on the right side of the figure. (B) 50 µg of cell lysate (as used in Fig. 2A) was resolved by 8% SDS-PAGE, followed by western transfer and probed with the anti-pp85 cortactin antibody. (C) By using the same lysates as in Fig. 2A, 30 µg of lysate was resolved by 8% SDS-PAGE and western blot analysis performed with rabbit anti-src antibodies to quantify the steady state levels of Src and the chimeric constructs (top panel), or with anti-phosphoY416 to detect the activation state of the Src or chimeric constructs.