Figure 7.
Overexpression of Cyclin B restores the mitotic index, cell cycle progression and reduces lagging chromosomes in TEM4-depleted cells. (A) Control and TEM4-depleted cells were transfected with GFP- or GFP-cyclin B for 48 h prior to total protein extraction with RIPA buffer. Western blotting was performed with the indicated antibodies. (B) Cells grown on coverslips were transfected as in A and synchronized in mitosis with 100 ng/ml of nocodazole for 12 h prior to fixation with PTEMF. Data show mean ± SEM, n = 3 independent experiments, N ∼ 200 cells. A two-way ANOVA test for significance was performed followed by Šídák’s multiple comparisons test to determine P values. (C) Mitotic progression of TEM4-depleted cells in the presence or absence of cyclin B. TEM4. Cells were transfected as in B and synchronized in mitosis after release from double thymidine block and the percentage of cells in each mitotic stage was calculated for all indicated conditions. Data shows mean ± SEM, n = 3 independent experiment, N ≥ 50 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. (D) Mitotic index in TEM4-depleted cells in the presence or absence of cyclin B. Cells were treated for 12 h with taxol. Data show means ± SEM, n = 3 independent experiments, N ≥ 25 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. (E) Quantifications of lagging chromosomes in TEM4 depleted in the presence or absence of cyclin B. Cells were treated for 12 h with 0.4 μΜ RO-3306 and released into fresh media for 1.5 h. Data show mean ± SEM, n = 3, N ≥ 25 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. Source data are available for this figure: SourceData F7. Refer to the image caption for details.

Overexpression of Cyclin B restores the mitotic index, cell cycle progression and reduces lagging chromosomes in TEM4-depleted cells. (A) Control and TEM4-depleted cells were transfected with GFP- or GFP-cyclin B for 48 h prior to total protein extraction with RIPA buffer. Western blotting was performed with the indicated antibodies. (B) Cells grown on coverslips were transfected as in A and synchronized in mitosis with 100 ng/ml of nocodazole for 12 h prior to fixation with PTEMF. Data show mean ± SEM, n = 3 independent experiments, N ∼ 200 cells. A two-way ANOVA test for significance was performed followed by Šídák’s multiple comparisons test to determine P values. (C) Mitotic progression of TEM4-depleted cells in the presence or absence of cyclin B. TEM4. Cells were transfected as in B and synchronized in mitosis after release from double thymidine block and the percentage of cells in each mitotic stage was calculated for all indicated conditions. Data shows mean ± SEM, n = 3 independent experiment, N ≥ 50 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. (D) Mitotic index in TEM4-depleted cells in the presence or absence of cyclin B. Cells were treated for 12 h with taxol. Data show means ± SEM, n = 3 independent experiments, N ≥ 25 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. (E) Quantifications of lagging chromosomes in TEM4 depleted in the presence or absence of cyclin B. Cells were treated for 12 h with 0.4 μΜ RO-3306 and released into fresh media for 1.5 h. Data show mean ± SEM, n = 3, N ≥ 25 cells per condition. A two-way ANOVA test for significance was performed followed by Tukey’s multiple comparisons test to determine P values. Source data are available for this figure: SourceData F7.

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