Overexpression of B⁰AT1 and TMEM27 in MDCK cells.

<p>A: immunofluorescence analysis of B⁰AT1-TMEM27 overexpressing MDCK cells. Apical co-localization (yellow) of B⁰AT1 (red) and TMEM27 (green) is visualized by confocal microscopy. Upper panels represent images taken parallel to the filter (x–y plane); lower panels show corresponding z–x reconstitutions. Bars = 5 µm. B: Apical uptake of L-Leu (100 µM, 10 min) in wild type (wt) or B⁰AT1 or B⁰AT1-TMEM27 overexpressing MDCK cells cultivated on filters. Data are represented as mean ± SEM (n = 3). Groups were compared by one-way ANOVA followed by Tukey post-test; * p≤0.05. C: MDCK cells overexpressing B⁰AT1-TMEM27 or EGFP. After viral transduction, cells were subcultured on plastic dishes for 5 or 10 times (passage 5 and 10, respectively) in standard cell culture medium. Western blotting experiments with antibodies directed against B⁰AT1, TMEM27 and EGFP were performed. Representative Western blotting images of 3 independent experiments are shown.