Fig. 2

Inhibition of NHEJ further decreases Fanca^−/− HSC renewal and repopulation. a Inhibition of DNA-PKcs further decreases Fanca^−/− HSPC proliferation. BM LSK cells isolated from WT or Fanca^−/− mice were treated with DNA-PKcs inhibitor NU7026 (10 μM) for 36 h and then plated in cytokine-supplemented methylcellulose medium. Colonies were enumerated on day 7 after plating. Results are means ± standard deviation (SD) of three independent experiments. *p < 0.05 or ***p < 0.001 vs WT vehicle control. b Inhibition of DNA-PKcs further compromises the repopulating capacity of Fanca^−/− HSPCs. BM LSK cells isolated from WT or Fanca^−/− mice were treated with DNA-PKcs inhibitor NU7026 (10 μM) for 36 h. Two thousand LSK cells, along with 200,000 c-Kit-depleted protector cells, were then transplanted into lethally irradiated BoyJ mice. The recipients were subjected to flow cytometric analysis for donor-derived LSK cells 16 weeks after BMT (n = 9–12 per group). *p < 0.05 or **p < 0.01 vs WT vehicle control. c Knockdown of Ku70 further decreases Fanca^−/− HSPC proliferation. BM LSK cells from WT or Fanca^−/− mice were transduced with lentiviruses co-expressing eGFP and scramble shRNA or shRNA targeting Ku70. Transduced cells were sorted for eGFP expression and then plated in cytokine-supplemented methylcellulose medium. Colonies were enumerated on day 7 after plating. Results are means ± standard deviation (SD) of three independent experiments. *p < 0.05, **p < 0.01, or ***p < 0.001 vs WT scramble shRNA control. d Knockdown of Ku70 compromises the repopulating capacity of Fanca^−/− HSPCs. Two thousand LSK cells described in c, along with 200,000 c-Kit-depleted protector cells, were then transplanted into lethally irradiated BoyJ mice. The recipients were subjected to flow cytometric analysis for donor-derived LSK cells 16 weeks after BMT (n = 9 per group). *p < 0.05 or ***p < 0.001 vs WT scramble shRNA control