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. 2004 Nov 24;101(49):17108–17113. doi: 10.1073/pnas.0407585101

Fig. 2.

Fig. 2.

Morphological and molecular analysis of Brca1Δ11/Δ11 MEF cells. (A and B) Morphology difference between WT (A) and Brca1Δ11/Δ11 (B) primary MEF cells in responding to nocodazole treatment at 24 h. Brca1Δ11/Δ11 MEF cells at mitotic phase exhibited significantly more fragmented (grape-like) cells. Phosphorylated histone H3 Ab staining also indicated that many mutant cells contained fragmented chromosomes. (C) Mitotic index determined by FACS analysis in primary MEF cells by using double staining with propidum iodide and an Ab to phosphorylated histone H3. (D and E) FACS assays of annexin V in WT and Brca1Δ11/Δ11 cells at 24 (D) and 36 (E) h after nocodazole treatment. (F and G) CDK1 kinase assay and cyclin B1 Western blot analysis of WT (F) and Brca1Δ11/Δ11 MEF (G) cells after nocodazole treatment at 0–60 h. CDK1 Western blotting to illustrate the equal amount of kinase was used. (H) Western blot analysis showing cyclin B1 levels in noninducible (29) and inducible (217) clones for Brca1 acute deletion. (I and J) Chromosome spreads showing premature sister-chromatid separation in Brca1 mutant (J) but not in control (I) cells.