Figure 9.

Wortmannin (WT) can agument changes of cell shape and spreading induced by Gas3/PMP22. (A) gas3/PMP22 FLAG was overexpressed in NIH3T3 cells by nuclear microinjection. After 6 h cells were treated or not with WT and LY294002; 12 h later cells were fixed and processed for immunofluorescence to detect Gas3/PMP22FLAG. Survival and morphological changes were scored as described in the text. Data represent arithmetic means ± SD for five independent experiments (altered shape, p < 0.001). (B) Immunofluorescence analysis of NIH3T3 cells expressing gas3/PMP22 in the presence of WT. NIH3T3 cells 24 h after seeding were microinjected with pEXV-gas3/PMP22FLAG (50 ng/μl). After 6 h cells were treated or not with WT; 12 h later cells were fixed and processed for immunofluorescence to visualize Gas3/PMP22FLAG. Bar, 25 μm. (C) AK and rhoA-V14 were coexpressed with gas3/PMP22 in NIH3T3 cells. After 6 h cells were treated or not with WT; 12 h later cells were fixed and processed for immunofluorescence to detect Gas3/PMP22FLAG. Survival and morphological changes were scored as described in the text. Data represent arithmetic means ± SD for five independent experiments (altered shape, p < 0.001).