Figure 7. Myo5B and Stx3 deletion results in selective cargo mislocalization. (A) Depletion of Stx3 (gStx3) or the expression of a dominant-negative Stx3 (mCitrine-Stx3_1-247) mutant does not affect the correct apical localization of DPPIV. Stx3_1-247 expression results in disrupted polarity (multilayer formation; Wiegerinck et al., 2014). (B) NHE3 is mislocalized from the brush border (highlighted by actin staining) to the cytoplasm in Stx3-depleted cells (gSTX3). (C) Correct localization of aminopeptidase N to the brush border (highlighted by actin staining) upon deletion of Myo5B. Arrowheads on the left mark the corresponding XY and XZ planes. Quantification and statistical analysis of indicated cargo/enzyme localization is shown to the right of corresponding confocal micrographs. Mean ± SD. ***, P < 0.005. n ≥ 100 cells. Bars, 10 µm. api, apical/brush border plane; mid, middle/nuclear plane.
Figure 7.

Myo5B and Stx3 deletion results in selective cargo mislocalization. (A) Depletion of Stx3 (gStx3) or the expression of a dominant-negative Stx3 (mCitrine-Stx3_1-247) mutant does not affect the correct apical localization of DPPIV. Stx3_1-247 expression results in disrupted polarity (multilayer formation; Wiegerinck et al., 2014). (B) NHE3 is mislocalized from the brush border (highlighted by actin staining) to the cytoplasm in Stx3-depleted cells (gSTX3). (C) Correct localization of aminopeptidase N to the brush border (highlighted by actin staining) upon deletion of Myo5B. Arrowheads on the left mark the corresponding XY and XZ planes. Quantification and statistical analysis of indicated cargo/enzyme localization is shown to the right of corresponding confocal micrographs. Mean ± SD. ***, P < 0.005. n ≥ 100 cells. Bars, 10 µm. api, apical/brush border plane; mid, middle/nuclear plane.

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