Figure 4. Ate1 −/− cells show an... | Rockefeller University Press

Figure 4.

$Ate1−/−cells show an increased association of MAP1S with microtubules. (A–C) Quantification of the LFQ intensity of MAP1S (A), MAP1A (B), and MAP1B (C) identified by mass spectrometry in microtubule preparations from Ate1+/+ and Ate1−/− cells. Mean ± SD, N = 3, Ate1+/+ versus Ate1−/−: *P = 0.016, two-tailed paired t test. Symbol colors indicate paired measurements. (D) Quantification of the RNA-level expression of Map1s in Ate1+/+ and Ate1−/− cells. Mean ± SD, N = 4, Wilcoxon matched-pairs signed rank test. Symbol colors indicate paired measurements. (E) Western blot for MAP1S (top, green) and tubulin (bottom, red) from Ate1+/+ and Ate1−/− cells. (F) Quantification of total MAP1S normalized to tubulin in Ate1+/+ and Ate1−/− cells. Mean ± SD, N = 3, two-tailed Wilcoxon matched-pairs signed rank test. Symbol colors indicate paired measurements. (G) Western blot for MAP1S (top, green) and tubulin (bottom, green) on soluble and polymerized tubulin fractions from Ate1+/+ and Ate1−/− cells. (H) Quantification of the fraction of MAP1S associated with the polymerized tubulin. Mean ± SEM, N = 3 (n = 12 total repeats per condition), Ate1+/+ versus Ate1−/−: *P = 0.0416, one-tailed paired t test. Symbol colors indicate paired average measurements. Violin plot represents the fraction of MAP1S in the pellet for individual repeats. (I) Immunofluorescence images of Ate1+/+ and Ate1−/− cells stained for MAP1S (green), ⍺-tubulin (magenta), and DAPI (blue). Scale bar = 20 μm. Tubulin and MAP1S are scaled to the same respective intensity as the wild-type image, for ease of visual comparison of intensity. (J) Western blot for total protein stain (left, red) and MAP1S (right, green) on immunoprecipitated MAP1S (IP) and input (In). Source data are available for this figure: SourceData F4. Refer to the image caption for details.$

Ate1 ^−/− cells show an increased association of MAP1S with microtubules. (A–C) Quantification of the LFQ intensity of MAP1S (A), MAP1A (B), and MAP1B (C) identified by mass spectrometry in microtubule preparations from Ate1^+/+ and Ate1^−/− cells. Mean ± SD, N = 3, Ate1^+/+ versus Ate1^−/−: *P = 0.016, two-tailed paired t test. Symbol colors indicate paired measurements. (D) Quantification of the RNA-level expression of Map1s in Ate1^+/+ and Ate1^−/− cells. Mean ± SD, N = 4, Wilcoxon matched-pairs signed rank test. Symbol colors indicate paired measurements. (E) Western blot for MAP1S (top, green) and tubulin (bottom, red) from Ate1^+/+ and Ate1^−/− cells. (F) Quantification of total MAP1S normalized to tubulin in Ate1^+/+ and Ate1^−/− cells. Mean ± SD, N = 3, two-tailed Wilcoxon matched-pairs signed rank test. Symbol colors indicate paired measurements. (G) Western blot for MAP1S (top, green) and tubulin (bottom, green) on soluble and polymerized tubulin fractions from Ate1^+/+ and Ate1^−/− cells. (H) Quantification of the fraction of MAP1S associated with the polymerized tubulin. Mean ± SEM, N = 3 (n = 12 total repeats per condition), Ate1^+/+ versus Ate1^−/−: *P = 0.0416, one-tailed paired t test. Symbol colors indicate paired average measurements. Violin plot represents the fraction of MAP1S in the pellet for individual repeats. (I) Immunofluorescence images of Ate1^+/+ and Ate1^−/− cells stained for MAP1S (green), ⍺-tubulin (magenta), and DAPI (blue). Scale bar = 20 μm. Tubulin and MAP1S are scaled to the same respective intensity as the wild-type image, for ease of visual comparison of intensity. (J) Western blot for total protein stain (left, red) and MAP1S (right, green) on immunoprecipitated MAP1S (IP) and input (In). Source data are available for this figure: SourceData F4.