Fig. 3: Detyrosinated α-tubulin accumulates on shrinking kinetochore microtubules.

a Immunofluorescence analysis of a 3D projection from a U2OS Parental cell using anti-tyrosinated (tyr) α-tubulin (white), anti-centromere antibody (ACA; magenta), anti-detyrosinated (detyr) α-tubulin antibodies (blue) and anti-EB1 (green); Grayscale panels show single channels for tyrosinated α-tubulin and detyrosinated α-tubulin (max intensity projection vs. sum projection). Scale bars 5 µm. a’ Single section from the same cell in (a); individual signals for each channel are also displayed in insets. b Line scan profile of the fluorescence intensities (F.I.) from the highlighted region from the inset in (a’). c Representation of the model of the distribution of tyrosinated and detyrosinated α-tubulin near oscillating kinetochores (kMTs = kinetochore microtubules). AP = anti-poleward, P = Poleward. d Quantification of the ratio between detyrosinated α-tubulin fluorescence intensity and tyrosinated α-tubulin fluorescence intensity of the growing (EB1 + ) kMTs and shrinking (EB1-) kMTs near oscillating kinetochores in the parental cell line; error bars represent mean and standard deviation; each point represents a kinetochore; quantifications from a pool of 3 independent experiments; at least 1 kinetochore pair per cell was measured (total kinetochore pairs = 95) from a total of 50 cells; exact p-values are displayed above each respective data set; significance p < 0.05, unpaired two-tailed t-test. e Quantification of the ratio of the detyrosinated α-tubulin normalized fluorescence intensity between the growing (EB1 + ) and shrinking (EB1-) kMTs from the same kinetochore pair, in the same data set as in (d). Source data are provided as a Source Data file.