Fig. 8. Overexpressed Gas3/PMP22 was route to the late endosomes. (A) NIH3T3 cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to LBPA. Arrowheads indicate vacuoles in the perinuclear region double positive for Gas3/PMP22 and LBPA. (B) NIH3T3 cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to LBPA. The arrow indicates large peripheral vacuoles negative for LBPA staining. (C) NIH3T3 cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to LBPA. Arrowheads indicate clusters of aggregated vacuoles in the perinuclear region double positive for Gas3/PMP22 and LBPA. (D) NIH3T3 cells were microinjected with Claudin-15-GFP. 15 hours later cells were fixed and probed with an antibody to LBPA. (E) NIH3T3 cells were microinjected with Claudin-15-GFP. 15 hours later cells were fixed and probed with an antibody to annexin II. (F) NIH3T3 cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to annexin II. Arrows indicate large peripheral vacuoles positive for annexin II staining. (G) NIH3T3 cells were microinjected with Gas3/PMP22-GFP. 15 hours later cells were fixed and probed with an antibody to LBPA and phalloidin TRITC. Arrows indicate large peripheral vacuoles coated with actin and arrowheads point to clusters of aggregated vacuoles in the perinuclear region double positive for Gas3/PMP22 and LBPA. Images were obtained using a Leica TCS confocal microscopy and in the overlay are displayed in pseudocolors: green (GFP fused proteins), red (LBPA, annexin II and actin) blue (LBPA). Bar, 5 µm.

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