MT regrowth assay in CSAP over-expressing cells.

Abstract

<p><b>(A, B, C)</b> MT regrowth assay of mitotic U2OS cells transiently over-expressing TrAP-centrin (a) or GFP-CSAP (b). After recovery from cold treatment for 0 min <b>(A)</b>, 3 min <b>(B)</b>, and 25 min <b>(C)</b>, the cells were stained for α-tubulin (Red), γ-tubulin (Green), and DNA (Blue). (c) Quantitative comparisons of α-tubulin are shown. Data represent the mean ± SD relative intensity. ** and *** indicate p < 0.01 and p < 0.005, respectively. <b>(D)</b> Mitotic U2OS cells transiently over-expressing TrAP-CSAP after cold treatment. Cells were stained for TrAP-CSAP (Green), γ-tubulin (Red), and DNA (Blue). <b>(E)</b> Images of mitotic U2OS cells transiently over-expressing GFP-Centrin (a) or GFP-CSAP (b) after 1 h of treatment with nocodazole before fixation. The cells were stained for α-tubulin (Red) and DNA (Blue). <b>(F, G)</b> Mitotic U2OS cells transiently over-expressing GFP-Centrin (a; Green) or GFP-CSAP (b) with nocodazole <b>(F)</b> or monastrol <b>(G)</b> for 12 h. Cells were stained for α-tubulin (Red) and DNA (Blue). Scale bar, 5 μm. <b>(H, I)</b> Quantitative comparisons of α-tubulin in cells treated with nocodazole <b>(H)</b> or monastrol <b>(I)</b> for 1 h. Data represent the mean ± SD relative intensity. *** indicates p < 0.005 (n = ~30).</p