Multimodal regulation of KT–MT dynamics by CLASP2. (A) Photoactivation experiments in live U2OS parental PA–GFP–α-tubulin cells and expressing the different mRFP-CLASP2γ constructs after CLASP RNAi. Panels represent DIC, the mRFP-CLASP2γ signal (red), and the PA–GFP–α-tubulin signal before photoactivation (Pre-PA), immediately after photoactivation (PA), and at 2 and 4.5 min after photoactivation. The PA–GFP–α-tubulin signal was inverted for better visualization. Scale bar is 5 µm. (B) Quantification of KT–MT half-life for all cell lines. The first column corresponds to the parental cell line and the remaining columns correspond to experiments performed after CLASP depletion. Each data point represents an individual cell. The boxes represent median and interquartile interval; the bars represent minimum and maximum values. Parental = 43 cells, pool of five independent experiments; Parental with RNAi = 20 cells, pool of six independent experiments; WT = 66 cells, pool of seven independent experiments; 2ea-3eeaa = 105 cells, pool of eight independent experiments; IP12 = 108 cells, pool of nine independent experiments; 2ea-IP12 = 52 cells, pool of 11 independent experiments; IP12-3eeaa = 55 cells, pool of four independent experiments; ΔC = 60 cells, pool of four independent experiments; ΔC-Gcn4 = 34 cells, pool of eight independent experiments; ΔC-Spc25 = 54 cells, pool of 12 independent experiments; C-term = 53 cells, pool of 12 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; Mann-Whitney U-Test. (C) Quantification of non-KT–MT half-life for all cell lines for the same dataset as in B. (D) Quantification of spindle poleward flux for all cell lines and same dataset as in B and C.