Fig. 1. Expression of the Gas3/PMP22-GFP. (A) Immunodetection of Gas3/PMP22-GFP in transfected U20S cells. U2OS cells were transfected with pEGFP-N1-Gas3/PMP22 and cellular extracts were treated with or without PNGase-F or Endo-H as indicated. Immunodetection was performed using the anti-GFP antibody. (B) Subcellular localization of Gas3/PMP22-GFP and Gas3/PMP22-L16P-GFP. Gas3/PMP22-GFP and Gas3/PMP22-L16PGFP were overexpressed in NIH3T3 by nuclear microinjection. 6 hours after microinjection, cells were fixed and double-stained to visualize GFP and ER, using biotinylated concanavalin A. Bar, 18 µm. (C) Time-lapse images of Schwann cells overexpressing Gas3/PMP22-GFP. Frames at selected hours after microinjection, as indicated, showing Schwann cells overexpressing Gas3/PMP22-GFP. Arrowheads underline a reduction in spreading; asterisks indicate floating dead cells. (D)Time-lapse images of Schwann cells overexpressing GFP. Frames at selected hours after microinjection showing Schwann cells overexpressing GFP. Bar, 30 µm.

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