Fig. 3. RhoB is involved in the intracellular targeting of Src but not of Yes and Fyn. (A) The colocalization of Myc-RhoB (upper panels) and Myc-RhoD (lower panels) with Src-WT, Yes-WT or Fyn-WT was studied in SYF^–/– cells stimulated with PDGF. Solid arrows indicate colocalization. (B) SYF^–/– cells expressing Fyn-WT-GFP and Myc-RhoB or Myc-RhoD were stimulated with PDGF and stained for activated SFK. (C) RhoB^+/+ (upper panels) or RhoB^–/– (lower panels) cells expressing Src-WT, Yes-WT or Fyn-WT were plated on fibronectin for 1 hour and stained for activated SFK. Solid arrows indicate localization of active protein at the cell periphery whereas broken arrows indicate inactive SFK retained in the perinuclear region. Localization of active SFK was detected using an anti-P-Y416-SFK antibody (Texas Red or Cy5 secondary), total protein using an anti-Src, anti-Yes or anti-Fyn antibody (FITC secondary) and Myc-Rho with an anti-Myc antibody (Texas Red secondary). Bars, 25 µm. Quantification shown in the Tables S1 and S2 in supplementary material.