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Figure 6


Fig. 6. EGF stimulation results in translocation of reggie-1 from plasma membrane into endosomes, and knockdown of reggie-1 interferes with EGF-induced actin remodeling. (A-F) Transfected HeLa cells were starved without serum for 16 hours and then stimulated with 100 ng/ml EGF for 15 minutes (B,D,F). Both wild-type R1-EGFP (B) and R1-myc (F) underwent an EGF-dependent translocation into an endosomal compartment, whereas the Y163F mutant (D) remained associated with the plasma membrane. (G,H) HeLa cells transfected with siRNA duplex oligoribonucleotides were starved for 16 hours and then stimulated with EGF for 5 minutes. Staining of the actin cytoskeleton with phalloidin revealed that EGF-induced changes in the actin cytoskeleton were impaired in reggie knockdown cells (H), which displayed fewer protrusions than the control cells (G). Panels A-D represent GFP fluorescence, whereas R1-myc was visualized in E,F by staining with an anti-myc antibody. Panels G,H show staining of the cells with fluorochrome-coupled phalloidin.





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