Fig. 5: PMCA4b promotes polarized vesicular trafficking.
From: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
a WGA uptake assay was performed on parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines. After 10 min incubation with WGA, cells were washed and transferred to a 37 °C incubator for 20 min. Dashed white squares indicate areas for magnification on (d). DAPI labels nuclei. b Yellow lines connect the center of the cell nucleus with WGA positive endocytic vesicles accumulated in the cytosol. c Intracellular distribution analysis of WGA positive vesicles. Red line indicates the distribution fit curve. Data were collected from 2 independent experiments, n(MCF-7) = 1214, n(sh-PMCA4) = 922, n(PMCA4b) = 1021, n(PMCA4bLA) = 1493 vesicles from 15 to 28 cells, and analyzed as described in the Methods section. d Statistical analysis of the distribution of WGA positive vesicles is illustrated by a box plot diagram with medians, interquartile range boxes and whiskers. Interquartile rage boxes represent the middle 50% of the data. Whiskers represent the ranges for the bottom 25% and the top 25% of the data values. Data were the same as (c) and analyzed with Levene’s test. P values: **P < 0.01, ***P < 0.001; non-significant difference is not labeled. e Magnification of the areas indicated by the dashed white squares on (a). Yellow arrows indicate colocalized WGA and PMCA4b positive vesicles.
