Fig. 1. EGF-receptor-dependent recruitment of SH2 domains to intracellular compartments containing internalized EGF. The tandem SH2 domains of PLC
1, fused to enhanced GFP (GFP-SH2), were used as a probe to visualize EGF receptor activation. GFP-SH2-transfected NR6 fibroblasts expressing the wild-type EGF receptor (NR6 WT) were either (A) untreated or (B) stimulated with Texas Red-conjugated EGF (EGF-TR) for 20 minutes. The live cells were subsequently visualized by confocal fluorescence microscopy as described in Materials and Methods. (C) GFP-SH2-transfected, EGF-TR-treated parental NR6 cells, which lack EGF receptors, were used as a control. Scale bar, 20 µm.
