Figure 5.
Depletion of MAP1S in Ate1−/−cells rescues decreased microtubule growth rate and increased microtubule stability. (A) Western blot showing MAP1S (top, green) and tubulin (bottom, red) in Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s. (B) Quantification of MAP1S normalized to tubulin in Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s. Mean ± SD, N = 5, two-tailed paired t-test. Symbol colors indicate paired measurements. (C) mCherry-EB3 signal (top) and tracks color-coded by mean straight-line speed ranging from 0 to 0.4 μm/s (bottom) from movies of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and overexpressing mCherry-EB3 (Video 5). Scale bar = 20 μm. (D) Quantification of EB3 comet velocity. Mean ± SEM, N = 3 (n = 15 cells per condition), Ate1+/+ + siCtrl versus Ate1−/− + siCtrl: ***P = 0.0003, Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: **P = 0.0014, Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ***P = 0.0003, one-way RM ANOVA corrected for multiple comparisons. Symbol colors indicate paired average measurements. Violin plot represents the distribution of average EB3 comet velocity for individual cells. (E) Immunofluorescence images of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and treated with 2 μM nocodazole for 0 or 10 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (F) Quantification of the average microtubule-positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 407–896 cells per condition), 10 min Ate1+/+ +siCtrl versus Ate1−/− + siCtrl: ****P < 0.0001, 10 min Ate1−/− + siCtrl versus Ate1+/+ + siMap1s: ****P < 0.0001, 10 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ****P < 0.0001, two-way RM ANOVA corrected for multiple comparisons. Symbol colors indicate paired average measurements. The truncated violin plot represents the distribution of the average microtubule-positive area for individual cells. Source data are available for this figure: SourceData F5. Refer to the image caption for details.

Depletion of MAP1S in Ate1 −/− cells rescues decreased microtubule growth rate and increased microtubule stability. (A) Western blot showing MAP1S (top, green) and tubulin (bottom, red) in Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s. (B) Quantification of MAP1S normalized to tubulin in Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s. Mean ± SD, N = 5, two-tailed paired t-test. Symbol colors indicate paired measurements. (C) mCherry-EB3 signal (top) and tracks color-coded by mean straight-line speed ranging from 0 to 0.4 μm/s (bottom) from movies of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and overexpressing mCherry-EB3 (Video 5). Scale bar = 20 μm. (D) Quantification of EB3 comet velocity. Mean ± SEM, N = 3 (n = 15 cells per condition), Ate1+/+ + siCtrl versus Ate1−/− + siCtrl: ***P = 0.0003, Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: **P = 0.0014, Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ***P = 0.0003, one-way RM ANOVA corrected for multiple comparisons. Symbol colors indicate paired average measurements. Violin plot represents the distribution of average EB3 comet velocity for individual cells. (E) Immunofluorescence images of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and treated with 2 μM nocodazole for 0 or 10 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (F) Quantification of the average microtubule-positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 407–896 cells per condition), 10 min Ate1+/+ +siCtrl versus Ate1−/− + siCtrl: ****P < 0.0001, 10 min Ate1−/− + siCtrl versus Ate1+/+ + siMap1s: ****P < 0.0001, 10 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ****P < 0.0001, two-way RM ANOVA corrected for multiple comparisons. Symbol colors indicate paired average measurements. The truncated violin plot represents the distribution of the average microtubule-positive area for individual cells. Source data are available for this figure: SourceData F5.

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