Figure 1.
Whole-body Gdf11 deletion does not alter frequency of fetal hematopoietic stem and progenitor cells (HSPCs) or affect hematopoietic reconstitution following FL transplantation. (A-C) Representative CD71/Ter-119 flow cytometry plots of EPCs isolated from E14.5 (A) Gdf11+/+, (B) Gdf11+/−, and (C) Gdf11−/− FLs. Cells previously gated as live, Lin−, CD41−. EPCs were identified by differential expression of CD71 and Ter-119, with cells progressing in maturity from S1 to S5.20 (D) Quantification of EPC frequency among live cells in E14.5 Gdf11+/+, Gdf11+/−, and Gdf11−/− FLs (n = 12 embryos per genotype; males and females pooled). (E-G) Representative CD48/CD150 flow cytometry plots of HSCs isolated from E14.5 (E) Gdf11+/+, (F) Gdf11+/−, and (G) Gdf11−/− FLs. Cells previously gated as live, Lin−, Sca-1+, c-Kit+. (H) Quantification of HSC frequency among live cells in E14.5 Gdf11+/+, Gdf11+/−, and Gdf11−/− FLs (n = 12 embryos per genotype; males and females pooled). (I) Experimental design. Gdf11+/+, Gdf11+/−, and Gdf11−/− embryos were harvested at E14.5. FL cells from sex- and genotype-matched embryos were pooled, and 1 × 106 FL cells were injected IV into lethally irradiated sex-matched CD45.1 recipients. Recipients were monitored for blood parameters and donor chimerism levels as indicated. (J) RBC counts, (K) hematocrit, and (L) hemoglobin levels in recipient mice at 0 (baseline), 1, 2, 3, and 4 weeks postirradiation and transplant (n = 20 recipients per donor genotype; males and females pooled). (M-P) Monthly peripheral blood CD45.2+ donor chimerism levels within (M) CD3+ T cells, (N) B220+ B cells, (O) CD11b+ Ly6G− monocytes, and (P) CD11b+ Ly6G+ neutrophils (n = 20 recipients per donor genotype; males and females pooled). (Q-R) CD45.2+ donor cell chimerism levels within BM (Q) Lin−Sca-1+c-Kit+ cells and (R) Lin−Sca-1+c-Kit+CD48−CD150+ HSCs (n = 20 recipients per donor genotype; males and females pooled). Data are plotted as (J-P) mean ± standard deviation (SD) or as (D, H, Q-R) individual data points overlaid with mean ± SD. Statistics for panels J-P were calculated using a repeated-measures ANOVA. Statistics for panels D, H, Q-R were calculated using a 1-way ANOVA with Bonferroni posttest correction. For all panels, no differences with P < .05 were detected. (D, H, Q-R) Circles: males; triangles: females. LSK, Lineage−Sca-1+c-Kit+.

Whole-body Gdf11 deletion does not alter frequency of fetal hematopoietic stem and progenitor cells (HSPCs) or affect hematopoietic reconstitution following FL transplantation. (A-C) Representative CD71/Ter-119 flow cytometry plots of EPCs isolated from E14.5 (A) Gdf11+/+, (B) Gdf11+/−, and (C) Gdf11−/− FLs. Cells previously gated as live, Lin−, CD41−. EPCs were identified by differential expression of CD71 and Ter-119, with cells progressing in maturity from S1 to S5.20  (D) Quantification of EPC frequency among live cells in E14.5 Gdf11+/+, Gdf11+/−, and Gdf11−/− FLs (n = 12 embryos per genotype; males and females pooled). (E-G) Representative CD48/CD150 flow cytometry plots of HSCs isolated from E14.5 (E) Gdf11+/+, (F) Gdf11+/−, and (G) Gdf11−/− FLs. Cells previously gated as live, Lin−, Sca-1+, c-Kit+. (H) Quantification of HSC frequency among live cells in E14.5 Gdf11+/+, Gdf11+/−, and Gdf11−/− FLs (n = 12 embryos per genotype; males and females pooled). (I) Experimental design. Gdf11+/+, Gdf11+/−, and Gdf11−/− embryos were harvested at E14.5. FL cells from sex- and genotype-matched embryos were pooled, and 1 × 106 FL cells were injected IV into lethally irradiated sex-matched CD45.1 recipients. Recipients were monitored for blood parameters and donor chimerism levels as indicated. (J) RBC counts, (K) hematocrit, and (L) hemoglobin levels in recipient mice at 0 (baseline), 1, 2, 3, and 4 weeks postirradiation and transplant (n = 20 recipients per donor genotype; males and females pooled). (M-P) Monthly peripheral blood CD45.2+ donor chimerism levels within (M) CD3+ T cells, (N) B220+ B cells, (O) CD11b+ Ly6G− monocytes, and (P) CD11b+ Ly6G+ neutrophils (n = 20 recipients per donor genotype; males and females pooled). (Q-R) CD45.2+ donor cell chimerism levels within BM (Q) Lin−Sca-1+c-Kit+ cells and (R) Lin−Sca-1+c-Kit+CD48−CD150+ HSCs (n = 20 recipients per donor genotype; males and females pooled). Data are plotted as (J-P) mean ± standard deviation (SD) or as (D, H, Q-R) individual data points overlaid with mean ± SD. Statistics for panels J-P were calculated using a repeated-measures ANOVA. Statistics for panels D, H, Q-R were calculated using a 1-way ANOVA with Bonferroni posttest correction. For all panels, no differences with P < .05 were detected. (D, H, Q-R) Circles: males; triangles: females. LSK, Lineage−Sca-1+c-Kit+.

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