Figure 2

PDGF receptors activate Abl kinase in hippocampal neurons. Acutely dissected hippocampal CA1 slices were incubated in the absence or presence of 10 ng/mL PDGF-BB for 10 minutes. A) Slices were homogenized and the lysate was incubated with anti-PDGFβ receptor antibodies precoupled to agarose beads. Total lysate and immunoprecipitated samples were immunoblotted with anti-Abl (top) or anti-PDGFβ receptor (bottom). Blots are representative of 7 experiments. B) Slices were homogenized and the lysates were immunoprecipitated with an anti-Abl antibody. Lysates and immunoprecipitates were resolved via SDS-Page electrophoresis and immunoblotted with anti-phospho-tyrosine (PY) or anti-Abl kinase antibodies. Blots are representative of 7 experiments. C) Anti-Src antibodies were used to immunoprecipitated Src after treatment of slices with 10 ng/mL PDGF-BB for 10 minutes. Immunoprecipitated samples were separated by SDS-electrophoresis and membranes were probed with anti-tyrosine 146 (Src-416), anti-tyrosine-527 (Src-527), or anti-Src (Src) antibodies. Blots are representative of 4–6 experiments. D) Lysates were immunoprecipitated with anti-Abl antibodies and membranes were probed with anti-PLCγ antibodies. Blot is representative of 5 experiments.