Figure 3.
Effects of kinesin-1 mutations implicated in autoinhibition on dynein-mediated early endosome transport. (A) Colony phenotypes of the kinA-GFP, kinA(1–894)-GFP, kinA∆IAK-GFP, kinAK895E-GFP, kinAE186K-GFP, kinAE178K,186K,K735R-GFP, and kinAE178K,P426L-GFP strains. (B) A quantitative analysis of colony diameter of the kinA-GFP (n = 19), kinA(1–894)-GFP (n = 19), kinA∆IAK-GFP (n = 25), kinAK895E-GFP (n = 22), kinAE186K-GFP (n = 19), kinAE178K,P426L-GFP (n = 22), and kinAE178K,186K,K735R-GFP (n = 20) strains. Ordinary one-way ANOVA (unpaired) with Tukey’s multiple comparisons test was used to analyze these data sets. Scatter plots with mean and SD values were generated by Prism 9. (C) Microscopic images showing the distributions of KinA-GFP and mCherry-RabA-labeled early endosomes (mCherry-RabA) in the kinA-GFP, kinA(1–894)-GFP, kinA∆IAK-GFP, and kinAK895E-GFP strains. Bar, 5 μm. (D) Line scans of KinA-GFP fluorescence intensity in the kinA-GFP (n = 31), kinA(1–894)-GFP (n = 61), kinA∆IAK-GFP (n = 61) and kinAK895E-GFP (n = 60) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. All values are relative to the peak mean value for kinA(1–894)-GFP, which is set as 1. The intensity of GFP near the hyphal tip (between 0 and 2.6 μm from hyphal tip) was significantly higher in the kinA(1–894)-GFP, kinA∆IAK-GFP or kinAK895E-GFP strain than in the kinA-GFP strain (P < 0.0001, two-way ANOVA with Tukey’s multiple comparisons test). (E) A quantitative analysis on the percentage of hyphal tips with the abnormal accumulation of early endosomes (Ordinary one-way ANOVA, unpaired). Three experiments were performed, and in each experiment, >80 hyphal tips were examined for each strain. Scatter plots with mean and SD values were generated by Prism 9. (F) Microscopic images showing the distributions of KinA-GFP and mCherry-RabA-labeled early endosomes (mCherry-RabA) in the kinAE186K-GFP, kinAE178K,186K,K735R-GFP, and kinAE178K,P426L-GFP strains. Bar, 5 μm. (G) Line scans of KinA-GFP fluorescence intensity in the kinAE186K-GFP (n = 30), kinAE178K,186K,K735R-GFP (n = 31), and kinAE178K,P426L-GFP (n = 30) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of GFP near the hyphal tip (between 0.065 and 1.885 μm from hyphal tip) was significantly different from each other among the three strains (P < 0.0001, two-way ANOVA with Tukey’s multiple comparisons test). (H) A quantitative analysis on the percentage of hyphal tips with the abnormal accumulation of early endosomes (Ordinary one-way ANOVA (unpaired) with Tukey’s multiple comparisons test). Three experiments were performed, and in each experiment, >40 hyphal tips were examined for each strain. Scatter plots with mean and SD values were generated by Prism 9.

Effects of kinesin-1 mutations implicated in autoinhibition on dynein-mediated early endosome transport. (A) Colony phenotypes of the kinA-GFP, kinA(1–894)-GFP, kinA∆IAK-GFP, kinAK895E-GFP, kinAE186K-GFP, kinAE178K,186K,K735R-GFP, and kinAE178K,P426L-GFP strains. (B) A quantitative analysis of colony diameter of the kinA-GFP (n = 19), kinA(1–894)-GFP (n = 19), kinA∆IAK-GFP (n = 25), kinAK895E-GFP (n = 22), kinAE186K-GFP (n = 19), kinAE178K,P426L-GFP (n = 22), and kinAE178K,186K,K735R-GFP (n = 20) strains. Ordinary one-way ANOVA (unpaired) with Tukey’s multiple comparisons test was used to analyze these data sets. Scatter plots with mean and SD values were generated by Prism 9. (C) Microscopic images showing the distributions of KinA-GFP and mCherry-RabA-labeled early endosomes (mCherry-RabA) in the kinA-GFP, kinA(1–894)-GFP, kinA∆IAK-GFP, and kinAK895E-GFP strains. Bar, 5 μm. (D) Line scans of KinA-GFP fluorescence intensity in the kinA-GFP (n = 31), kinA(1–894)-GFP (n = 61), kinA∆IAK-GFP (n = 61) and kinAK895E-GFP (n = 60) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. All values are relative to the peak mean value for kinA(1–894)-GFP, which is set as 1. The intensity of GFP near the hyphal tip (between 0 and 2.6 μm from hyphal tip) was significantly higher in the kinA(1–894)-GFP, kinA∆IAK-GFP or kinAK895E-GFP strain than in the kinA-GFP strain (P < 0.0001, two-way ANOVA with Tukey’s multiple comparisons test). (E) A quantitative analysis on the percentage of hyphal tips with the abnormal accumulation of early endosomes (Ordinary one-way ANOVA, unpaired). Three experiments were performed, and in each experiment, >80 hyphal tips were examined for each strain. Scatter plots with mean and SD values were generated by Prism 9. (F) Microscopic images showing the distributions of KinA-GFP and mCherry-RabA-labeled early endosomes (mCherry-RabA) in the kinAE186K-GFP, kinAE178K,186K,K735R-GFP, and kinAE178K,P426L-GFP strains. Bar, 5 μm. (G) Line scans of KinA-GFP fluorescence intensity in the kinAE186K-GFP (n = 30), kinAE178K,186K,K735R-GFP (n = 31), and kinAE178K,P426L-GFP (n = 30) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of GFP near the hyphal tip (between 0.065 and 1.885 μm from hyphal tip) was significantly different from each other among the three strains (P < 0.0001, two-way ANOVA with Tukey’s multiple comparisons test). (H) A quantitative analysis on the percentage of hyphal tips with the abnormal accumulation of early endosomes (Ordinary one-way ANOVA (unpaired) with Tukey’s multiple comparisons test). Three experiments were performed, and in each experiment, >40 hyphal tips were examined for each strain. Scatter plots with mean and SD values were generated by Prism 9.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal