Fig. 7. Colocalization of endogenous ezrin with the 5-HT_4(a) but not the 5-HT_4(b) splice variant in the presence of NHERF in NIH-3T3 cells. Cells were transfected by electroporation with plasmids encoding either HA-NHERF-RhoTag-5-HT_4(a)R/pIRES2 (A, top, B) or the HA-NHERF-RhoTag-5-HT_4(b)R/pIRES2 (A, bottom, C) construct. They were permeabilized and examined by confocal microscopy. Left, medium and right panels illustrate endogenous ezrin staining (anti-ezrin antibody 1:1,000), RhoTag staining (anti-RhoTag antibody 1:100) and merged images, respectively. Mid-nuclear slices (inlay: zoom) and vertical scans in the z axis (z) (along the black lines in the x-y scans) are depicted. Representative cells are represented. Bars, 10 µm.