Figure 4.
Hemin impaired erythropoiesis in vivo via induction of IFNα. (A) Plasma IFNα levels of various mice as indicated (N = 6). (B-C) Representative western blot and quantitative analyses of Stat2 levels in the enriched BM erythroid cells from PBS- or hemin-injected mice (N = 3). (D) Effects of IFNα injection to AA mice on BM BFU-E colonies, CFU-E colonies, and erythroblasts (N = 6). (E-F) Representative western blot and quantitative analyses of Stat2 levels in the enriched BM erythroid cells from PBS- or IFNα-injected AA mice (N = 3). (G) Quantitative analysis of pStat5 levels as assessed by flow cytometry in BM erythroid cells of PBS- or IFNα-injected AA mice. (H-I) Representative western blot and quantitative analyses of pErk levels in enriched BM erythroid cells from PBS- or IFNα-injected AA mice. The fold changes of pErk were normalized to the pErk level from BM erythroid cells of PBS-injected AA mice stimulated with EPO at 1 U/mL (N = 3). (J-K) Representative western blot and quantitative analyses of pStat5 levels in the enriched BM erythroid cells of AA mice treated with IFNα in vitro at indicated concentrations and then stimulated with EPO at 0 or 1 U/mL. The fold changes were normalized to the pStat5 level in the enriched erythroid cells without IFNα treatment and stimulated with EPO at 1 U/mL (N = 3). (L) Quantitative analyses of Cish messenger RNA (mRNA) levels in enriched BM erythroid cells from AA mice injected with PBS or IFNα (N = 3). (M-N) Representative western blot and quantitative analyses of Stat2 levels in enriched BM erythroid cells from AA and SS mice (N = 3). (O) Quantitative analyses of Cish mRNA levels in enriched BM erythroid cells of AA and SS mice (N = 3). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.

Hemin impaired erythropoiesis in vivo via induction of IFNα. (A) Plasma IFNα levels of various mice as indicated (N = 6). (B-C) Representative western blot and quantitative analyses of Stat2 levels in the enriched BM erythroid cells from PBS- or hemin-injected mice (N = 3). (D) Effects of IFNα injection to AA mice on BM BFU-E colonies, CFU-E colonies, and erythroblasts (N = 6). (E-F) Representative western blot and quantitative analyses of Stat2 levels in the enriched BM erythroid cells from PBS- or IFNα-injected AA mice (N = 3). (G) Quantitative analysis of pStat5 levels as assessed by flow cytometry in BM erythroid cells of PBS- or IFNα-injected AA mice. (H-I) Representative western blot and quantitative analyses of pErk levels in enriched BM erythroid cells from PBS- or IFNα-injected AA mice. The fold changes of pErk were normalized to the pErk level from BM erythroid cells of PBS-injected AA mice stimulated with EPO at 1 U/mL (N = 3). (J-K) Representative western blot and quantitative analyses of pStat5 levels in the enriched BM erythroid cells of AA mice treated with IFNα in vitro at indicated concentrations and then stimulated with EPO at 0 or 1 U/mL. The fold changes were normalized to the pStat5 level in the enriched erythroid cells without IFNα treatment and stimulated with EPO at 1 U/mL (N = 3). (L) Quantitative analyses of Cish messenger RNA (mRNA) levels in enriched BM erythroid cells from AA mice injected with PBS or IFNα (N = 3). (M-N) Representative western blot and quantitative analyses of Stat2 levels in enriched BM erythroid cells from AA and SS mice (N = 3). (O) Quantitative analyses of Cish mRNA levels in enriched BM erythroid cells of AA and SS mice (N = 3). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.

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