Fig. 6. Gas3/PMP22-induced vacuoles are PIP₂-positive and trap MHC I. (A) U2OS cells were transfected with Gas3/PMP22-VSV and PHGFP or with Gas3/PMP22-L16P-VSV and PH-GFP. 40 hours later cells were fixed and probed with an antibody to VSV. Bar, 8 µm. (B) NIH3T3 cells were co-injected with Gas3/PMP22-VSV and PH-GFP, with Gas3/PMP22-L16P-VSV and PH-GFP or with Gas3/PMP22-MG-VSV and PH-GFP. 18 hours later cells were fixed and probed with antibody to VSV to visualize Gas3/PMP22. Bar, 8 µm. (C) IMR90-E1A cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to MHC I. Bar, 4 µm. (D) IMR90-E1A cells were microinjected with Gas3/PMP22-GFP. 15 hours later they were allowed to internalize MHC I antibody for 60 minutes and then were fixed and processed for visualizing internalized antibody. Bar, 5 µm. (E) U2OS cells were microinjected with Gas3/PMP22-GFP. 18 hours later they were allowed to internalize MHC I antibody for 60 minutes and then were fixed and processed for visualizing internalized antibody. Bar, 5 µm.