Figure 6: Deficiency of 14-3-3γ decreases the surface expression of ANO1 in U251 cells.

(a) Representative traces of A01-sensitive currents in U251 cells transfected with Sc shRNA, 14-3-3γ shRNA-1, and/or a GFP-14-3-3γ-insensitive form of 14-3-3γ. (b) Summary bar graph showing normalized A01-sensitive current density as in (a) at +80 mV. Data are expressed as mean ± SEM; ***P < 0.001. (c) Cell surface biotinylation results from membrane protein fractions from U251 cells infected with adenovirus containing Sc shRNA or 14-3-3γ shRNA-1. Western blot images were cropped for clarification. Full blot images were included in supplementary information. (d) A normalized bar graph showing data obtained from three independent experiments as in (c). Data are expressed as mean ± SEM; *P < 0.05. (e) Representative immunocytochemical images of U251 cells transfected with Sc shRNA or 14-3-3γ shRNA-1. Scale bar, 10 μm. (f) Pearson’s correlation coefficients denoting covariance of fluorescence signals were calculated using Nikon software as in (e). Data are expressed as mean ± SEM; *P < 0.05.