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.