Figure S4. EHBP-1 sequesters exocytic... | Rockefeller University Press

Figure S4.

$EHBP-1 sequesters exocytic carriers through a mechanism dependent on RAB-10. (A–A″) Confocal images showing the subcellular localization of GFP-RAB-10. Statistical data is shown as box-and-whisker plots with 10th–90th percentile (n = 24 cells from eight animals of each genotype; dots, outliers; boundaries, quartiles; two-tailed Mann–Whitney test). (B) Western blotting showing the results of membrane fractionation for GFP-RAB-10. S: supernatant; P: pellet. (C and C′) Confocal images showing colocalization between RAB-10 and mitochondria. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; two-tailed Mann-Whitney test). (D and D′) Confocal images showing colocalization between GFP or mCherry fusion proteins. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; one-way ANOVA test with Dunn’s multiple comparison). (E and E′) The images were acquired using a TIRF microscope showing liposomes captured by slides immobilized with StrepII-EHBP-1(ΔC2). Statistical data is shown as box-and-whisker plots with the 10th–90th percentile (n = 24 areas; dots, outliers; boundaries, quartiles; two-way ANOVA with Bonferroni post-test). (F and F′) Confocal images showing colocalization between hTAC-GFP and Mito-mCherry or Mito-EHBP-16AALAA-mCherry or Mito-mCherry-RAB-10. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; one-way ANOVA test with Dunn’s multiple comparison). Source data are available for this figure: SourceData FS4. Refer to the image caption for details.$

EHBP-1 sequesters exocytic carriers through a mechanism dependent on RAB-10. (A–A″) Confocal images showing the subcellular localization of GFP-RAB-10. Statistical data is shown as box-and-whisker plots with 10th–90th percentile (n = 24 cells from eight animals of each genotype; dots, outliers; boundaries, quartiles; two-tailed Mann–Whitney test). (B) Western blotting showing the results of membrane fractionation for GFP-RAB-10. S: supernatant; P: pellet. (C and C′) Confocal images showing colocalization between RAB-10 and mitochondria. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; two-tailed Mann-Whitney test). (D and D′) Confocal images showing colocalization between GFP or mCherry fusion proteins. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; one-way ANOVA test with Dunn’s multiple comparison). (E and E′) The images were acquired using a TIRF microscope showing liposomes captured by slides immobilized with StrepII-EHBP-1(ΔC2). Statistical data is shown as box-and-whisker plots with the 10th–90th percentile (n = 24 areas; dots, outliers; boundaries, quartiles; two-way ANOVA with Bonferroni post-test). (F and F′) Confocal images showing colocalization between hTAC-GFP and Mito-mCherry or Mito-EHBP-1^6AALAA-mCherry or Mito-mCherry-RAB-10. The statistical analysis of colocalization was calculated as the Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; one-way ANOVA test with Dunn’s multiple comparison). Source data are available for this figure: SourceData FS4.