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. 2009 Apr 9;28(10):1442–1452. doi: 10.1038/emboj.2009.96

Figure 3.

Figure 3

Interdependence between the Ska complex. (A) HeLa S3 cells were treated for 48 h with control (Gl2) and Ska1, Ska2 and Ska3-specific siRNAs, respectively. Mitotic cells from Gl2-treated cells arrested with nocodazole or Ska1, Ska2 and Ska3-depleted cells were collected by shake-off. Equal amounts of cell extracts were separated by SDS–PAGE and probed by western blotting with anti-Ska3, anti-Ska1, anti-Ska2 and anti-α-Tubulin antibodies. (B) Bar graph of corresponding intensity profile for western signal of (A), normalized against signal for anti-α-Tubulin. (C) Asynchronously growing cells were treated for 48 h with control (Gl2) and Ska1, Ska2 and Ska3-specific siRNAs, respectively. The mitotic indices were determined by light microscopy. Histograms show averages from four independent experiments. Error bars represent standard deviation (s.d.). (D) Cells were treated for 48 h with control (Gl2), Ska1- and Ska3-specific siRNAs, respectively, then fixed with PTEMF and stained with anti-Ska3 (left panels) or anti-Ska1 (right panels) antibodies (red), CREST serum (far red, depicted in green) or anti-α-Tubulin (green). DNA was visualized using DAPI (blue). Bar=10 μm.