Figure 5.
Effect of PARP-2 and PARP-1 deficiency on Eμ-Myc BM pre-B cell proliferation, cell cycle, and apoptosis. (A-B) In vivo BM pre-B cell proliferation was determined by intraperitoneal injection of 5-week-old mice of the indicated genotypes with BrdU (1 mg/6 g mouse weight). BM cells were harvested at 2 hours after the onset of injection, and BrdU incorporation on pre-B cells was analyzed by flow cytometry. Representative histograms (A) from 2 independent experiments including ≥ 2 mice from each genotype are shown. Numbers indicate percentage of proliferating (BrdU+) cells. (B) Bars represent mean ± SEM (SEM) values of the percentage of BrdU+ cells. (C) Representative flow cytometric dot plots showing the cell cycle status of BM pre-B cells in mice from the indicated genotypes. BM pre-B cells were stained with Ki67 to identify cycling cells and 4′,6-diamidino-2-phenylindole to measure DNA content. The percentage of cells in each quadrant represents the mean from ≥ 6 mice in each group. (D) Graph showing the percentage of BM pre-B cells for each genotype that are in G0, G1, S, and G2/M phases of the cell cycle. (E-F) Large-scale gene expression and GSEA showed a significant enrichment for G2/M checkpoint (E) and intrinsic apoptotic signaling (F) pathways in pre-B cells from Eμ-MycT/+Parp-2−/− mice compared with Eμ-MycT/+ wild-type (WT) control cells. At the left is the GSEA enrichment plot; at the right, the normalized gene expression heatmap of genes involved in that pathways. (G) Representative dot plots showing active caspase-3 staining in BM pre-B cells for each genotype. (H) Bars represent the percentage of cells positive for active caspase-3. Values represent mean ± SEM obtained from ≥ 6 mice per genotype. (I) Quantitative reverse transcription polymerase chain reaction analysis in BM pre-B cells of genes involved in cell cycle and apoptosis. Samples were normalized according to β-actin expression levels. Results are expressed as fold expression compared with levels measured in WT cells. Values represent mean ± SEM obtained from 3 independent experiments. *P < .05, **P < .01, ***P < .001.

Effect of PARP-2 and PARP-1 deficiency on Eμ-Myc BM pre-B cell proliferation, cell cycle, and apoptosis. (A-B) In vivo BM pre-B cell proliferation was determined by intraperitoneal injection of 5-week-old mice of the indicated genotypes with BrdU (1 mg/6 g mouse weight). BM cells were harvested at 2 hours after the onset of injection, and BrdU incorporation on pre-B cells was analyzed by flow cytometry. Representative histograms (A) from 2 independent experiments including ≥ 2 mice from each genotype are shown. Numbers indicate percentage of proliferating (BrdU+) cells. (B) Bars represent mean ± SEM (SEM) values of the percentage of BrdU+ cells. (C) Representative flow cytometric dot plots showing the cell cycle status of BM pre-B cells in mice from the indicated genotypes. BM pre-B cells were stained with Ki67 to identify cycling cells and 4′,6-diamidino-2-phenylindole to measure DNA content. The percentage of cells in each quadrant represents the mean from ≥ 6 mice in each group. (D) Graph showing the percentage of BM pre-B cells for each genotype that are in G0, G1, S, and G2/M phases of the cell cycle. (E-F) Large-scale gene expression and GSEA showed a significant enrichment for G2/M checkpoint (E) and intrinsic apoptotic signaling (F) pathways in pre-B cells from Eμ-MycT/+Parp-2−/− mice compared with Eμ-MycT/+ wild-type (WT) control cells. At the left is the GSEA enrichment plot; at the right, the normalized gene expression heatmap of genes involved in that pathways. (G) Representative dot plots showing active caspase-3 staining in BM pre-B cells for each genotype. (H) Bars represent the percentage of cells positive for active caspase-3. Values represent mean ± SEM obtained from ≥ 6 mice per genotype. (I) Quantitative reverse transcription polymerase chain reaction analysis in BM pre-B cells of genes involved in cell cycle and apoptosis. Samples were normalized according to β-actin expression levels. Results are expressed as fold expression compared with levels measured in WT cells. Values represent mean ± SEM obtained from 3 independent experiments. *P < .05, **P < .01, ***P < .001.

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