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Figure 3


Fig. 3. PLAP is mislocalized in fully polarized FRT cells stably lacking TI-VAMP. (A) Confocal Z and X-Y sections (top and bottom) of FRT cells stably expressing PLAP, and FRT cells stably expressing si TI-VAMP::PLAP, grown on filters for 4 days and labeled with antibody against PLAP and secondary antibodies coupled to FITC. PLAP is mislocalized to the lateral membrane of FRT si TI-VAMP::PLAP (right panel). (B) FRT PLAP and FRT si TI-VAMP::PLAP cells were grown on filters for 4 days and labeled with LC-biotin (Biot) added to the apical (Ap) or basolateral (Bl) surface. After lysis, PLAP was immunoprecipitated with a specific antibody, run on SDS-PAGE and revealed with HRP-streptavidin (lane one and panel three). One tenth of the immunoprecipitate (Tot) was run on SDS-PAGE and assayed with an antibody against PLAP (panels two and four). Bars, 10 µm. IgG, immunoglobulin G.





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