Integrity of the Pericentriolar Material Is Essential for Maintaining Centriole Association during M Phase
Fig 7
Effects of nocodazole and BI2536 on PCM dispersal in STLC-treated cells.
(A) HeLa cells were cultured in the presence of STLC for 20 h. Nocodazole was added at the 9th hour point. The cells were immunostained with the α-tubulin (green) and γ-tubulin (red) antibodies. The cells were also immunostained with centrin-2 and CEP135 antibodies to determine centriole association. Scale bar, 10 μm. (B, C) HeLa cells were treated with STLC for 20 h. Nocodazole (200 ng/ml) or BI2536 (200 nM) was added at 9th h. The cells were immunostained with antibodies specific to pericentrin, CEP192, CEP215 and γ-tubulin (green) along with centrin-2 (red). Scale bar, 10 μm. (C) The γ-tubulin staining patterns were categorized as discrete and dispersed. The experiments were repeated twice, with over 150 cells at each group. Values are means and standard deviations. (D) HeLa cells were arrested at S phase with thymidine and released in the presence of STLC. At the indicated time points, the cells were coimmunostained with antibodies specific to CEP135 and γ-tubulin to determine centriole association and PCM dispersal, respectively. The experiments were repeated twice, with over 100 cells at each group. Values are means and standard deviations. (E) HeLa cells were treated with STLC for 9 h and then with nocodazole for 11 h. The cells were then transferred to a nocodazole-free medium and cultured for up to 4 h. At the indicated time points, the cells were coimmunostainined with antibodies specific to centrin-2 and pericentrin. DNA was stained with DAPI.