Figure 2.
KinA(1–894)-GFP fusion proteins form a prominent accumulation near the microtubule plus ends. (A) Colony phenotypes of the strains containing kinA-GFP and kinA(1–894)-GFP in comparison to wild type and the kinAK895* mutant. (B) A quantitative analysis of colony diameter of the wild type (n = 24), kinAK895* (n = 24), kinA-GFP (n = 24) and kinA(1–894)-GFP (n = 23) strains (Kruskal–Wallis ANOVA test (unpaired) with Dunn’s multiple comparisons test). Scatter plots with mean and SD values were generated by Prism 9. (C) Localization of KinA-GFP or KinA(1–894)-GFP and mCherry-RabA-labeled early endosomes in strains containing one of the GFP fusions. Hyphal tip is indicated by a yellow arrowhead. Bar, 5 μm. (D) Line scans of KinA-GFP fluorescence intensity in the kinA-GFP (n = 33 hyphae) and kinA(1–894)-GFP (n = 32 hyphae) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The GFP intensity was significantly higher in the kinA(1–894)-GFP strain than in the kinA-GFP strain near the hyphal tip (between 0 and 3.51 μm from hyphal tip; P < 0.0001, two-way ANOVA with Bonferroni’s multiple comparisons test). (E) The hyphal-tip localization of KinA(1–894)-GFP in the alcA-nudAHC conditional-null mutant. Images were taken from cells grown on glucose, a repressive medium for the regulatable alcA promoter, which allows the expression of the nudA gene (encoding dynein heavy chain) to be turned off. Hyphal tip is indicated by a yellow arrowhead. Bar, 5 μm. (F) Line scans of KinA-GFP fluorescence intensity in the kinA(1–894)-GFP strain (n = 20 hyphae) and the alcA-nudAHC, kinA(1–894)-GFP strain (n = 20). XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of KinA-GFP between 0 and 11 μm from hyphal tip was not significantly different in these two strains (P > 0.9999, two-way ANOVA with Bonferroni’s multiple comparisons test). (G) Line scans of mCherry-RabA (early endosomes) fluorescence intensity in the kinA-GFP (n = 33 hyphae) and kinA(1–894)-GFP (n = 32 hyphae) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of mCherry-RabA near the hyphal tip (between 0.715 and 3.12 μm from hyphal tip) was significantly higher in the kinA(1–894)-GFP strain than in the kinA-GFP strain (P < 0.0001, two-way ANOVA with Bonferroni’s multiple comparisons test).

KinA (1–894) -GFP fusion proteins form a prominent accumulation near the microtubule plus ends. (A) Colony phenotypes of the strains containing kinA-GFP and kinA(1–894)-GFP in comparison to wild type and the kinAK895* mutant. (B) A quantitative analysis of colony diameter of the wild type (n = 24), kinAK895* (n = 24), kinA-GFP (n = 24) and kinA(1–894)-GFP (n = 23) strains (Kruskal–Wallis ANOVA test (unpaired) with Dunn’s multiple comparisons test). Scatter plots with mean and SD values were generated by Prism 9. (C) Localization of KinA-GFP or KinA(1–894)-GFP and mCherry-RabA-labeled early endosomes in strains containing one of the GFP fusions. Hyphal tip is indicated by a yellow arrowhead. Bar, 5 μm. (D) Line scans of KinA-GFP fluorescence intensity in the kinA-GFP (n = 33 hyphae) and kinA(1–894)-GFP (n = 32 hyphae) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The GFP intensity was significantly higher in the kinA(1–894)-GFP strain than in the kinA-GFP strain near the hyphal tip (between 0 and 3.51 μm from hyphal tip; P < 0.0001, two-way ANOVA with Bonferroni’s multiple comparisons test). (E) The hyphal-tip localization of KinA(1–894)-GFP in the alcA-nudAHC conditional-null mutant. Images were taken from cells grown on glucose, a repressive medium for the regulatable alcA promoter, which allows the expression of the nudA gene (encoding dynein heavy chain) to be turned off. Hyphal tip is indicated by a yellow arrowhead. Bar, 5 μm. (F) Line scans of KinA-GFP fluorescence intensity in the kinA(1–894)-GFP strain (n = 20 hyphae) and the alcA-nudAHC, kinA(1–894)-GFP strain (n = 20). XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of KinA-GFP between 0 and 11 μm from hyphal tip was not significantly different in these two strains (P > 0.9999, two-way ANOVA with Bonferroni’s multiple comparisons test). (G) Line scans of mCherry-RabA (early endosomes) fluorescence intensity in the kinA-GFP (n = 33 hyphae) and kinA(1–894)-GFP (n = 32 hyphae) strains. XY graphs with mean (solid lines) and SEM (shading) were generated by Prism 9. The intensity of mCherry-RabA near the hyphal tip (between 0.715 and 3.12 μm from hyphal tip) was significantly higher in the kinA(1–894)-GFP strain than in the kinA-GFP strain (P < 0.0001, two-way ANOVA with Bonferroni’s multiple comparisons test).

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