Figure 3.
Loss of EHBP-1 leads to increased dynamics in RAB-10-labeled exocytic carriers. (A and A′) Confocal images showing the morphology of endogenous VIT-2-associated endosome (VIT-2-mNeonGreen-3xFlagKI) in different genetic backgrounds. The data are shown as box-and-whisker plots with 10th–90th percentile (n = 24 cells from eight animals of each genotype; dots, outliers; boundaries, quartiles; one-way ANOVA test with Dunn’s multiple comparison). (B and B′) Volume confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI, mCherry-RAB-10, and EHBP-1-BFP. Line scan profiles of the three channels of the white line were conducted using ImageJ. (C and C′) Confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI and wrmScarlet-3xFlag-RAB-10KI in different genetic backgrounds. (D and D′) Confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI and EHBP-1-mCherrySC in different genetic backgrounds. Statistical analysis of colocalization was calculated as 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). (E–E″) Confocal images showing the morphology of endogenous RAB-10-associated endosome (mNeonGreen-3xFlag-RAB-10KI). 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). (F) Spanning disk confocal images showing the dynamics of GFP-RAB-10-labeled endosomes. Red arrowheads represent the movement of RAB-10-associated structures toward neighboring structures, resulting in eventual convergence. Green arrowheads illustrate fission events within RAB-10-associated structures. The moving routes were tracked using TrackMate. The tiny rings indicate the endosomal structures, while the zigzag lines indicate the dynamic trajectory of these structures. Refer to the image caption for details.

Loss of EHBP-1 leads to increased dynamics in RAB-10-labeled exocytic carriers. (A and A′) Confocal images showing the morphology of endogenous VIT-2-associated endosome (VIT-2-mNeonGreen-3xFlagKI) in different genetic backgrounds. The data are shown as box-and-whisker plots with 10th–90th percentile (n = 24 cells from eight animals of each genotype; dots, outliers; boundaries, quartiles; one-way ANOVA test with Dunn’s multiple comparison). (B and B′) Volume confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI, mCherry-RAB-10, and EHBP-1-BFP. Line scan profiles of the three channels of the white line were conducted using ImageJ. (C and C′) Confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI and wrmScarlet-3xFlag-RAB-10KI in different genetic backgrounds. (D and D′) Confocal images showing colocalization between VIT-2-mNeonGreen-3xFlagKI and EHBP-1-mCherrySC in different genetic backgrounds. Statistical analysis of colocalization was calculated as 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). (E–E″) Confocal images showing the morphology of endogenous RAB-10-associated endosome (mNeonGreen-3xFlag-RAB-10KI). 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). (F) Spanning disk confocal images showing the dynamics of GFP-RAB-10-labeled endosomes. Red arrowheads represent the movement of RAB-10-associated structures toward neighboring structures, resulting in eventual convergence. Green arrowheads illustrate fission events within RAB-10-associated structures. The moving routes were tracked using TrackMate. The tiny rings indicate the endosomal structures, while the zigzag lines indicate the dynamic trajectory of these structures.

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