Subcellular localizations of HRG-1, HCP1, and DMT1A-I in polarized or non-polarized MDCK cells. A. GFP-tagged HRG-1 (a) or mCherry-tagged HCP1 (b) were cotransfected into HEp-2 cells. The cells were cultured on glass coverslips for 48 h after replating. B. MDCK cells stably expressing GFP-tagged HRG-1 (a and d) and mCherry-tagged HCP1 (b) or mCherry-tagged DMT1A-I (e) were grown on Transwell and analyzed after their polarization. C. Cell surface labeling assay. MDCK cells stably expressing GFP-tagged HRG-1, mCherry-tagged HCP1, or mCherry-tagged DMT1A-I were grown on Transwell, and each apical or basolateral surface proteins were biotinylated. The biotinylated proteins were immunoprecipitated with protein A conjugated with anti-GFP or anti-mCherry polyclonal antibodies, and analyzed by Western blot. Similar results were obtained in 3 independent experiments.
Yanatori et al. BMC Cell Biology 2010 11:39 doi:10.1186/1471-2121-11-39