SDPN-1 directs the localization of RTKN-1 in recycling endosomes. (A) Western blot showing GST pulldown with in vitro–translated HA-tagged AMPH-1 and SDPN-1. (B) Coimmunoprecipitation assay analyzing RTKN-1–SDPN-1 interaction. (C) Western blot showing GST pulldown with in vitro–translated HA-SDPN-1. (D and D′) Confocal image showing colocalization between SDPN-1-GFP and mCherry-tagged RTKN-1 or RTKN-1ΔPRM in the intestinal cells. The white arrowheads indicate structures labeled by both GFP and mCherry. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (E and E′) Confocal images showing the distribution of RTKN-1-GFP. Box-and-whisker plots (n = 18 cells): 10–90th percentile; dots, outliers; red midline, median of WT; boundaries, quartiles. ***, P < 0.001 by one-way ANOVA followed by Dunn’s post hoc multiple comparison test. (F and F′) Confocal image showing colocalization between GFP-RME-1 and mCherry-tagged RTKN-1 or RTKN-1ΔPRM in the intestinal cells. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (G and G′) Confocal image showing colocalization between GFP-Utrophin-CH and Tubby (Tb)-PH(R332H)-mCherry in the intestinal cells. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (H and H′) Confocal images showing the distribution of hTAC-GFP. Box-and-whisker plots (n = 18 cells): 10–90th percentile; dots, outliers; red midline, median of WT; boundaries, quartiles. **, P < 0.01; ***, P < 0.001 by one-way ANOVA followed by Dunn’s post hoc multiple comparison test. The black asterisks in the image panels indicate intestinal lumen. Scale bars, 10 µm. IB, immunoblot; IP, immunoprecipitation; MC, mCherry.
Figure 10.

SDPN-1 directs the localization of RTKN-1 in recycling endosomes. (A) Western blot showing GST pulldown with in vitro–translated HA-tagged AMPH-1 and SDPN-1. (B) Coimmunoprecipitation assay analyzing RTKN-1–SDPN-1 interaction. (C) Western blot showing GST pulldown with in vitro–translated HA-SDPN-1. (D and D′) Confocal image showing colocalization between SDPN-1-GFP and mCherry-tagged RTKN-1 or RTKN-1ΔPRM in the intestinal cells. The white arrowheads indicate structures labeled by both GFP and mCherry. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (E and E′) Confocal images showing the distribution of RTKN-1-GFP. Box-and-whisker plots (n = 18 cells): 10–90th percentile; dots, outliers; red midline, median of WT; boundaries, quartiles. ***, P < 0.001 by one-way ANOVA followed by Dunn’s post hoc multiple comparison test. (F and F′) Confocal image showing colocalization between GFP-RME-1 and mCherry-tagged RTKN-1 or RTKN-1ΔPRM in the intestinal cells. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (G and G′) Confocal image showing colocalization between GFP-Utrophin-CH and Tubby (Tb)-PH(R332H)-mCherry in the intestinal cells. Pearson’s correlation coefficients for GFP and mCherry signals are calculated; error bar is 95% CI (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (H and H′) Confocal images showing the distribution of hTAC-GFP. Box-and-whisker plots (n = 18 cells): 10–90th percentile; dots, outliers; red midline, median of WT; boundaries, quartiles. **, P < 0.01; ***, P < 0.001 by one-way ANOVA followed by Dunn’s post hoc multiple comparison test. The black asterisks in the image panels indicate intestinal lumen. Scale bars, 10 µm. IB, immunoblot; IP, immunoprecipitation; MC, mCherry.

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