Figure S3.
Ate1−/−cells show increased microtubule stability, and knockdown of Map1s can rescue microtubule stability in Ate1−/−cells. (A) Immunofluorescence images of Ate1+/+, Ate1−/−, Ate1−/− + Ate1-GFP cells treated with 2 μM nocodazole for 0, 30, or 60 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (B) Quantification of the microtubule area per cell before nocodazole treatment. Mean ± SEM, N = 6 (n = 1,217–1,635 cells per condition), one-way RM ANOVA corrected for multiple comparisons. (C) Quantification of the average microtubule positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 650–1,137 cells per condition), 30 min Ate1+/+ versus Ate1−/−: ****P < 0.0001, 30 min Ate1−/− versus Ate1−/− + Ate1-GFP: ****P < 0.0001, 60 min Ate1+/+ versus Ate1−/−: ***P = 0.0009, 60 min Ate1−/− versus Ate1−/− + Ate1-GFP: ***P = 0.0002, two-way RM ANOVA corrected for multiple comparisons. For (B and C), symbol colors indicate paired average measurements, and truncated violin plots represent the distribution of average microtubule-positive area for individual cells. (D) Immunofluorescence images of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and treated with 2 μM nocodazole for 0, 30, or 60 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (E) Quantification of the microtubule area per cell before nocodazole treatment. Mean ± SEM, N = 6 (n = 1,263–2,042 cells per condition), one-way RM ANOVA corrected for multiple comparisons. (F) Quantification of the average microtubule-positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 769–1,174 cells per condition), 30 min Ate1+/+ + siCtrl versus Ate1−/− + siCtrl: ****P < 0.0001, 30 min Ate1+/+ + siCtrl versus Ate1−/− + siMap1s: *P = 0.0123, 30 min Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: ****P < 0.0001, 30 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ****P < 0.0001, 60 min Ate1+/+ +siCtrl versus Ate1−/− + siCtrl: ***P = 0.001, 60 min Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: ***P = 0.0005, 60 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ***P = 0.0001, two-way RM ANOVA corrected for multiple comparisons. For (E and F), symbol colors indicate paired average measurements, and truncated violin plots represent the distribution of average microtubule-positive area for individual cells. Refer to the image caption for details.

Ate1 −/− cells show increased microtubule stability, and knockdown of Map1s can rescue microtubule stability in Ate1 −/− cells. (A) Immunofluorescence images of Ate1+/+, Ate1−/−, Ate1−/− + Ate1-GFP cells treated with 2 μM nocodazole for 0, 30, or 60 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (B) Quantification of the microtubule area per cell before nocodazole treatment. Mean ± SEM, N = 6 (n = 1,217–1,635 cells per condition), one-way RM ANOVA corrected for multiple comparisons. (C) Quantification of the average microtubule positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 650–1,137 cells per condition), 30 min Ate1+/+ versus Ate1−/−: ****P < 0.0001, 30 min Ate1−/− versus Ate1−/− + Ate1-GFP: ****P < 0.0001, 60 min Ate1+/+ versus Ate1−/−: ***P = 0.0009, 60 min Ate1−/− versus Ate1−/− + Ate1-GFP: ***P = 0.0002, two-way RM ANOVA corrected for multiple comparisons. For (B and C), symbol colors indicate paired average measurements, and truncated violin plots represent the distribution of average microtubule-positive area for individual cells. (D) Immunofluorescence images of Ate1+/+ and Ate1−/− cells transfected with siCtrl or siMap1s and treated with 2 μM nocodazole for 0, 30, or 60 min, extracted, and stained for ⍺-tubulin (magenta) and DAPI (blue). Scale bar = 10 μm. (E) Quantification of the microtubule area per cell before nocodazole treatment. Mean ± SEM, N = 6 (n = 1,263–2,042 cells per condition), one-way RM ANOVA corrected for multiple comparisons. (F) Quantification of the average microtubule-positive area per cell normalized to t = 0. Mean ± SEM, N = 3 (n = 769–1,174 cells per condition), 30 min Ate1+/+ + siCtrl versus Ate1−/− + siCtrl: ****P < 0.0001, 30 min Ate1+/+ + siCtrl versus Ate1−/− + siMap1s: *P = 0.0123, 30 min Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: ****P < 0.0001, 30 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ****P < 0.0001, 60 min Ate1+/+ +siCtrl versus Ate1−/− + siCtrl: ***P = 0.001, 60 min Ate1−/− + siCtrl versus Ate1+/+ +siMap1s: ***P = 0.0005, 60 min Ate1−/− + siCtrl versus Ate1−/− + siMap1s: ***P = 0.0001, two-way RM ANOVA corrected for multiple comparisons. For (E and F), symbol colors indicate paired average measurements, and truncated violin plots represent the distribution of average microtubule-positive area for individual cells.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal