Intracellular localization of GFP-tagged MICA mutant molecules lacking
the cytoplasmic domain in polarized MDCK cells. (A) MDCK
cells were transiently transfected with the pEGFP-MICAdel plasmid
(which lacks the 47 residues C terminus of the protein). Confocal
fluorescence micrographs (x–y plane), beginning at the
apical membrane (section 1, Upper Left) and ending at
the basal membrane (section 9, Lower Right) were
acquired in 1.0-μm increments. Arrow in optical section 4 indicates
the plane of vertical section. (B) Confocal fluorescence
micrograph (x–z plane) shows the distribution of
GFP-tagged MICA mutant molecules lacking the cytoplasmic domain along
the apical–basal axis. GFP-tagged mutant MICA-derived fluorescence is
colored by green and nuclei by blue. AM, apical membrane; BM, basal
membrane. The scale bar is 10 μm. (C) Immunoblot
analysis of transiently expressed GFP-tagged mutant MICA molecules in
MDCK cells. MDCK cells, transiently transfected with the pEGFP-MICA
(MICA-GFP) or with pEGFP-MICAdel (MICAdel-GFP) plasmids, were
immunoblotted with anti-MICA mouse serum. A control consisted of MDCK
cells stably expressing GFP molecules alone. Sizes of marker proteins
used in kDa are shown on the left.