Fig. 1. Src, Fyn and Yes are distinctly spatially regulated. (A) SYF^–/– cells expressing Src-WT (wild-type Src), Yes-WT or Fyn-WT were maintained in serum-free medium (upper panels) or stimulated with PDGF (lower panels). Localization of active SFK was detected using an anti-P-Y416-SFK antibody (Texas Red secondary) and total protein using an anti-Src, anti-Yes or anti-Fyn antibody (FITC secondary). Solid arrows indicate SFK at the cell periphery whereas broken arrows indicate SFKs at the perinuclear region of the cell. Bars, 25 µm. (B) Western blot analysis of SYF^–/– cells expressing Src-WT (left panels), Yes-WT (middle panels) or Fyn-WT (right panels) in serum-free medium (–) or stimulated with PDGF. Activation was detected using an anti-P-Y416-SFK antibody (lower panels). Quantification shown in Table S1 in supplementary material.