Fig. 9. Gas3/PMP22 requires Arf6 to induce membrane internalization. (A) Arf6, Arf6-Q67L and Arf6-T27N were co-expressed with Gas3/PMP22-GFP in NIH3T3 cells. After 18 hours from microinjection cells were fixed and processed for immunofluorescence to visualize Gas3/PMP22-GFP and Arf6. Cells were scored for accumulation of vacuoles double positive for Arf6 and Gas3/PMP22. Data represent arithmetic means±s.d. for four independent experiments. (B) Arf6, Arf6-Q67L and Arf6-T27N were co-expressed with PH-GFP in NIH3T3 cells. After 18 hours from microinjection cells were fixed and processed for immunofluorescence to visualize PH-GFP and Arf6. Cells were scored for accumulation of vacuoles. Data represent arithmetic means±s.d. for four independent experiments. (C) Arf6 and Arf6-T27N were co-expressed with Gas3/PMP22-GFP in NIH3T3 cells. After 18 hours from microinjection cells were fixed and processed for immunofluorescence to visualize Gas3/PMP22-GFP and LBPA. Cells were scored for accumulation vesicles double positive for Gas3/PMP22 and LBPA. Data represent arithmetic means ± s.d. for four independent experiments.