FIG. 6.
The Y697F mutation of M-CSFR results in altered M-CSFR and Gab3 phosphorylation. (A) Effects of Y697F mutation on Gab3 phosphorylation. FD/Fms WT/Gab3V5/Mona and FD/Fms Y697F/Gab3V5/Mona cells were stimulated with 2,500 U of M-CSF per ml at 37°C for the indicated times. Proteins in cell lysates were immunoprecipitated (IP) using anti-Gab3 antibody. Immunoprecipitates were separated by SDS-7.5% PAGE and visualized by immunoblotting (IB) using antiphosphotyrosine (αpY; 1:10,000) or anti-Gab3 (1:100) antibody. (B) Effects of Y697F mutation on M-CSFR phosphorylation. FD/Fms WT/Gab3V5 and FD/Fms Y697F/Gab3V5 cells were stimulated with M-CSF as described above, and proteins in cell lysates were immunoprecipitated using anti-M-CSFR antibody. Immunoprecipitates were separated by SDS-7.5% PAGE and visualized by immunoblotting using antiphosphotyrosine (1:10,000) or anti-M-CSFR (1:500) antibody.