|
TECPR2 regulates membrane localization of SNX17 on early endosomes. A Lysates of HeLa cells treated with indicated siRNA and immunoblotted (IB) with indicated antibodies. The signal of mature Itgβ1 is marked with an arrowhead, and the lower band indicates the precursor Itgβ1 form. Densitometric analysis of mature Itgβ1, normalized to the control siRNA-treated sample, is shown. B Representative confocal micrographs of HeLa cells treated with the indicated siRNA, followed by immunostaining with anti-Rab5 and anti-SNX17 antibodies. Arrowheads in the insets indicate colocalized pixels. Scale bars: 10 µm (main); 5 µm (inset). C Quantification of SNX17 puncta number per cell from control and TECPR2-depleted HeLa cells. The values are represented as described for Fig. 2B. The error bar represents mean ± SD from three independent experiments (****p < 0.0001; unpaired two-tailed Student’s t-test). D Quantification of the Pearson’s colocalization coefficient (PCC) of SNX17 with Rab5 in HeLa cells treated with the indicated siRNA. The values are represented as described for Fig. 2B. The error bar represents mean ± SD from three independent experiments (****p < 0.0001; unpaired two-tailed Student’s t-test). E Live-cell imaging of HeLa cells treated with the indicated siRNA and expressing GFP-SNX17 (green). Before imaging, antibody-based labeling of surface Itgβ1 was performed, and cells were incubated in complete media at 37 °C to allow endocytosis of Itgβ1 (see Supplementary Movie 8). Arrowheads indicate endosomes positive for both GFP-SNX17 and endocytosed Itgβ1. Scale bar: 5 µm. F Quantification of the PCC of GFP-SNX17 with endocytosed Itgβ1 in HeLa cells treated with indicated siRNA. The values are represented as described for Fig. 2B. The error bar represents mean ± SD from three independent experiments (****p < 0.0001; unpaired two-tailed Student’s t-test). Source data are provided as a Source Data file.
|
