RTKN-1 impedes UNC-60A–mediated actin disassembly. (A and A′) Live cell fluorescence images showing the dynamic correlation between GFP-Utrophin-CH– and hTAC-GFP–labeled structures. White arrowheads indicate the hTAC-labeled mobile membrane carrier or dynamic tubules. Pearson's correlation coefficients were calculated; error bars are 95% CIs (n = 12 animals). ***, P < 0.001 by Mann-Whitney test. (B and B′) The actin-binding potential of GST-RTKN-1 was measured using the cosedimentation assay. Multiple concentrations were deployed to make a curve to estimate interaction (n = 3 independent experiments). (C) Actin polymerization was not affected by the presence of GST-RTKN-1 at various concentrations. (D) F-actin depolymerization was not affected by the presence of GST-RTKN-1 at various concentrations. (E) GST-RTKN-1 deferred GST-UNC-60A–mediated F-actin depolymerization, t_1/2 has been indicated. (F and F′) Confocal images showing the colocalization between endogenous UNC-60A and Utrophin-CH–labeled actin structures. OE, overexpression. Mander's coefficients were calculated; error bars are 95% CIs (n = 12 animals). **, P < 0.01; ***, P < 0.001 by Mann-Whitney test. (F′′) Fluorescence intensity measurement of GFP-Utrophin-CH. 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. (G and G′) Confocal images showing the colocalization between endogenous UNC-60A and RTKN-1-GFP. Pearson's correlation coefficients were calculated; error bars are 95% CIs (n = 12 animals). Scale bars, 10 µm. P, pellet; S, supernatant.