Figure 8.
TNF-α treatment prevents loss of H3H9me3 HSC function in vivo independently of their level of DNA damage. (A and B) Percentage of GFP-negative donor contribution in blood in mice transplanted with NIR, IR, or IR + TNF-α cells at 7 (A) and 14 wk (B) after reconstitution. (C and D) LSK GFP− (C) or LSK CD34−Flk2−CD48− (D) GFP-negative donor HSC contribution in the BM 14 wk after reconstitution. One-way ANOVA Tukey’s multiple comparison test. (E) Percentage of GFP-negative donor contribution in blood in mice secondary transplanted with pool of NIR, IR, or IR + TNF-α mice from the primary reconstitution. One-way ANOVA Tukey’s multiple comparison test. (F) Experimental design for TNF-α treatment after IR in vivo and reconstitution experiments using HSCs sorted from mice 1 mo after TBI and treated with TNF-α 6 h after IR (IR + TNFα 6 h), treated with TNF 1 and 13 h after IR (IR + TNFα 1–13 h), irradiated but non-treated with TNF-α (IR), or non irradiated and non treated (NIR); BMMC, bone marrow mononuclear cells. (G and H) Percentage of CD45.2 donor contribution in blood in mice transplanted with NIR, IR, or IR + TNF-α cells at 6 (G) and 11 wk (H). (I and J) Percentage of LSK−CD45.2 (I) or LSK−CD34−Flk2−CD45.2+ (J) donor HSC contribution in the BM 11 wk after reconstitution. One-way ANOVA Tukey’s (I) or or Dunnett’s (J) multiple comparison test. (K and L) γH2AX foci number 30 min and 24 h after IR in vitro with or without prior TNF-α treatment (K) or 1 mo after IR in vivo with or without TNF-α treatment before IR (IR + TNF-α) or after IR (IR + TNF-α 1–13 h; L). One-way ANOVA Tukey’s multiple comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

TNF-α treatment prevents loss of H3H9me3 HSC function in vivo independently of their level of DNA damage. (A and B) Percentage of GFP-negative donor contribution in blood in mice transplanted with NIR, IR, or IR + TNF-α cells at 7 (A) and 14 wk (B) after reconstitution. (C and D) LSK GFP− (C) or LSK CD34−Flk2−CD48− (D) GFP-negative donor HSC contribution in the BM 14 wk after reconstitution. One-way ANOVA Tukey’s multiple comparison test. (E) Percentage of GFP-negative donor contribution in blood in mice secondary transplanted with pool of NIR, IR, or IR + TNF-α mice from the primary reconstitution. One-way ANOVA Tukey’s multiple comparison test. (F) Experimental design for TNF-α treatment after IR in vivo and reconstitution experiments using HSCs sorted from mice 1 mo after TBI and treated with TNF-α 6 h after IR (IR + TNFα 6 h), treated with TNF 1 and 13 h after IR (IR + TNFα 1–13 h), irradiated but non-treated with TNF-α (IR), or non irradiated and non treated (NIR); BMMC, bone marrow mononuclear cells. (G and H) Percentage of CD45.2 donor contribution in blood in mice transplanted with NIR, IR, or IR + TNF-α cells at 6 (G) and 11 wk (H). (I and J) Percentage of LSK−CD45.2 (I) or LSK−CD34−Flk2−CD45.2+ (J) donor HSC contribution in the BM 11 wk after reconstitution. One-way ANOVA Tukey’s (I) or or Dunnett’s (J) multiple comparison test. (K and L) γH2AX foci number 30 min and 24 h after IR in vitro with or without prior TNF-α treatment (K) or 1 mo after IR in vivo with or without TNF-α treatment before IR (IR + TNF-α) or after IR (IR + TNF-α 1–13 h; L). One-way ANOVA Tukey’s multiple comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

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