Figure 7.
Figure 7. Chk2 is required for spindle checkpoint signaling in unperturbed early prometaphase. (A) Phosphorylated Aurora B–S331 (pS331). Cells were transfected with negative siRNA (control) or Chk2 siRNA (siChk2). (B) Cyclin B fluorescence in cells transfected as in A. ***, P < 0.001. A minimum of 20 cells per treatment was analyzed. Error bars show SDs. (C) Cells expressing Mad2-GFP were treated as in A. (D) Cells expressing Mps1-GFP were transfected as in A and treated with 2 µM AZ3146 for 1 h. (E) Tetracycline-induced CHO WT, S331A, or S331E Aurora B cells expressing Mps1-GFP were treated with 2 µM AZ3146 for 1 h. (F) WT or S331A Aurora B cells expressing Mad2-GFP were induced with tetracycline. Boxed values show mean green/red fluorescence intensity ± SDs. Values in square brackets show kinetochore pairs and number of cells analyzed. Bars, 5 µm. The insets in A, C, and D–F show 1.7× magnification of kinetochores.

Chk2 is required for spindle checkpoint signaling in unperturbed early prometaphase. (A) Phosphorylated Aurora B–S331 (pS331). Cells were transfected with negative siRNA (control) or Chk2 siRNA (siChk2). (B) Cyclin B fluorescence in cells transfected as in A. ***, P < 0.001. A minimum of 20 cells per treatment was analyzed. Error bars show SDs. (C) Cells expressing Mad2-GFP were treated as in A. (D) Cells expressing Mps1-GFP were transfected as in A and treated with 2 µM AZ3146 for 1 h. (E) Tetracycline-induced CHO WT, S331A, or S331E Aurora B cells expressing Mps1-GFP were treated with 2 µM AZ3146 for 1 h. (F) WT or S331A Aurora B cells expressing Mad2-GFP were induced with tetracycline. Boxed values show mean green/red fluorescence intensity ± SDs. Values in square brackets show kinetochore pairs and number of cells analyzed. Bars, 5 µm. The insets in A, C, and D–F show 1.7× magnification of kinetochores.

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