![Figure 2. Phenotypic analysis of MSPCs and introduction of WT FANCG gene restores normal MSPC proliferation and survival. (A) Representative expression of surface markers on WT and Fancg−/− MSPCs from 4 individual groups of cells was evaluated by flow cytometric analysis. Isotype controls are shown in dashed lines, and experimental samples are shown in gray area. (B) [3H]Thymidine incorporation in WT and Fancg−/− MSPCs was measured after stimulation with DMEM containing 0% to 10% FBS. Data are expressed as mean (± SE) from 4 individual experiments (*P < .001 for Fancg−/− vs WT cells). (C) At baseline and 24 and 48 hours after FBS depletion, the percentage of annexin/PI+ cells was analyzed by flow cytometry. Results of 4 independent experiments are shown. *P < .01 at 24 hours; **P < .001 at 48 hours for comparison of WT with Fancg−/− MSPCs. (D) Representative photographs of WT and Fancg−/− MSPC cultures after β-galactosidase staining are shown. Senescent cells were light blue cytoplasmic cells as pointed by red arrows. (E) Quantitative summary showed *P < .001 for Fancg−/− versus WT senescent cells from 3 independent experiments. (F) Flow cytometric analysis shows the percent enhanced green fluorescent protein (EGFP) expression on the MSPCs from WT mice (left) and Fancg−/− mice (right). (G) After transduction, cell proliferation was evaluated by counting the cell numbers in each culture condition, and data are shown as the fold change of cell numbers over the basal level in each genotype of cells after stimulation with DMEM containing 5% FBS (mean ± SE) from 4 individual experiments. *P < .01 for Fancg−/− with vector control versus WT cells or Fancg−/− cells transduced with WT FANCG sequence. (H) After transduction, cell apoptosis was evaluated by flow cytometry after deprivation of FBS up to 48 hours, and data are expressed as percentage of annexin V on different genotypes of MSPCs (mean ± SE) from 4 individual experiments. *P < .01 for Fancg−/− with vector control versus WT cells or Fancg−/− cells transduced with WT FANCG sequence.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/113/10/10.1182_blood-2008-07-168138/4/zh8013093263002f.jpeg?Expires=1722655395&Signature=dOCqAA7eVy9zLDswVmopOt7ZwEz6JtRC3rM5DVovL9DbsVVSJRhs9Pp8uCY5I4eipmbjIgICKT7mb8b4e0FwM5V182g-Ennk8r2GQ-sVM5mZaPt5FkP2sx-2aSThtrTugdZ5~zaRw7CJhdsHpQqx9e-39rgIrpz34MCgPTFqWsYrJJp~ggmmdCA5T4QQEO6Hobi49qxiBROTvp~w~Jiq745cTS6l5ByGbfYAJxWUMUV7aShcaSI8dYvUisRM8kJ4NiIvjLwa8s4fvfSWfnH15gDW93kip5zfTYax6nYfrFt46MpUf~bIRt2MDYjcnR30hYlX7TOrztnv4aLVlkQQpQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Phenotypic analysis of MSPCs and introduction of WT FANCG gene restores normal MSPC proliferation and survival. (A) Representative expression of surface markers on WT and Fancg−/− MSPCs from 4 individual groups of cells was evaluated by flow cytometric analysis. Isotype controls are shown in dashed lines, and experimental samples are shown in gray area. (B) [3H]Thymidine incorporation in WT and Fancg−/− MSPCs was measured after stimulation with DMEM containing 0% to 10% FBS. Data are expressed as mean (± SE) from 4 individual experiments (*P < .001 for Fancg−/− vs WT cells). (C) At baseline and 24 and 48 hours after FBS depletion, the percentage of annexin/PI+ cells was analyzed by flow cytometry. Results of 4 independent experiments are shown. *P < .01 at 24 hours; **P < .001 at 48 hours for comparison of WT with Fancg−/− MSPCs. (D) Representative photographs of WT and Fancg−/− MSPC cultures after β-galactosidase staining are shown. Senescent cells were light blue cytoplasmic cells as pointed by red arrows. (E) Quantitative summary showed *P < .001 for Fancg−/− versus WT senescent cells from 3 independent experiments. (F) Flow cytometric analysis shows the percent enhanced green fluorescent protein (EGFP) expression on the MSPCs from WT mice (left) and Fancg−/− mice (right). (G) After transduction, cell proliferation was evaluated by counting the cell numbers in each culture condition, and data are shown as the fold change of cell numbers over the basal level in each genotype of cells after stimulation with DMEM containing 5% FBS (mean ± SE) from 4 individual experiments. *P < .01 for Fancg−/− with vector control versus WT cells or Fancg−/− cells transduced with WT FANCG sequence. (H) After transduction, cell apoptosis was evaluated by flow cytometry after deprivation of FBS up to 48 hours, and data are expressed as percentage of annexin V on different genotypes of MSPCs (mean ± SE) from 4 individual experiments. *P < .01 for Fancg−/− with vector control versus WT cells or Fancg−/− cells transduced with WT FANCG sequence.
