Figure 4
Figure 4. Iex-1 is necessary and sufficient for TPO-mediated DSB repair in HSPCs. (A) Percentage of γH2AX-positive LSK cells isolated from nontreated WT and Iex-1−/− mice. Data are means + SEM (n = 3). (B) Kinetics of γH2AX foci resolution after IR (2 Gy) of WT and Iex-1−/− LSK cells cultured in complete medium (Comp) or without TPO. (C) WT LSK cells were infected with lentiviruses expressing GFP only or GFP with Iex-1-WT or Iex-1-ΔFTF and treated as in (B). For (B) and (C), the results represent means + SEM of 3 independent experiments performed with pools of 5 to 10 mice. (D) WT and Iex-1−/− mice were treated with TPO (8 µg/kg) or phosphate-buffered saline 30 minutes before TBI (2 Gy). γH2AX staining in LSK-CD34− cells isolated 16 hours after TBI. (E-F) γH2AX foci (E) and comet tail resolution (F) after IR of WT and Mpl−/− LSK cells infected as in (C) and cultured in the presence of TPO complete medium. Means + SEM are shown. (E) n = 4. (F) Data are means + SEM of tail moment values analyzed in at least 35 cells.

Iex-1 is necessary and sufficient for TPO-mediated DSB repair in HSPCs. (A) Percentage of γH2AX-positive LSK cells isolated from nontreated WT and Iex-1−/− mice. Data are means + SEM (n = 3). (B) Kinetics of γH2AX foci resolution after IR (2 Gy) of WT and Iex-1−/− LSK cells cultured in complete medium (Comp) or without TPO. (C) WT LSK cells were infected with lentiviruses expressing GFP only or GFP with Iex-1-WT or Iex-1-ΔFTF and treated as in (B). For (B) and (C), the results represent means + SEM of 3 independent experiments performed with pools of 5 to 10 mice. (D) WT and Iex-1−/− mice were treated with TPO (8 µg/kg) or phosphate-buffered saline 30 minutes before TBI (2 Gy). γH2AX staining in LSK-CD34 cells isolated 16 hours after TBI. (E-F) γH2AX foci (E) and comet tail resolution (F) after IR of WT and Mpl−/− LSK cells infected as in (C) and cultured in the presence of TPO complete medium. Means + SEM are shown. (E) n = 4. (F) Data are means + SEM of tail moment values analyzed in at least 35 cells.

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