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Fig. 3. Cellular localization of Pyk2, ArgBP2 and Cbl in PC12 cells. (A) PC12 cells stably expressing PDGF ß-receptors were grown on collagen I-coated coverslips and subjected to immunofluorescence with anti-Pyk2 (N-19), anti-Cbl antibodies (C-15) and anti-ArgBP2 antibodies. In the upper panel, Pyk2 is visualized in green by Alexa 488-labeled secondary donkey anti-goat antibodies (left), Cbl (middle) and ArgBP2 (right) are stained red with TRITC-labeled swine anti-rabbit secondary antibodies. In the lower panel, F-actin is stained blue by Alexa 305-labeled phalloidin (left), the merged picture between Pyk2, Cbl and F-actin is in the middle and the superimposed image of ArgBP2 and F-actin is on the right. Left and middle panels are from one experiment and the right panel from a separate experiment. (B) PC12 cells stably expressing PDGF ß-receptors were grown on collagen I-coated coverslips and subjected to immunofluorescence after 48 hours of PDGF-BB (50 ng/ml) stimulation. F-actin is stained with Alexa 305-labeled phalloidin (upper panel) or FITC-phalloidin (lower panel). Immunofluorescence was performed using goat polyclonal anti-Pyk2 (N-19), rabbit polyclonal anti-Cbl (C-15) and anti-ArgBP2 as indicated. Pyk2 was visualized by Alexa 488-labeled donkey anti-goat antibodies and Cbl (upper panel) and ArgBP2 (lower panel) by TRITC-labeled swine anti-rabbit antibodies. Upper and lower panels are form two different experiments.
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