Figure 1.
Conditional deletion of TGFβ1 in Mks increases apoptosis. (A) White blood cell (WBC), RBC, and platelet (PLT) counts are shown for TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 16 per group). (B) Marrow (n = 12 per group) and spleen (n = 5 per group) cellularity are shown for TGFβ1FL/FL (gray) and TGFβ1ΔMk/ΔMk mice (black). (C) The number of Lin− (those not expressing mature lineage markers CD3, B220, CD11b, Gr1, or Ter119) marrow cells per leg (femur and tibia) is shown (n = 13 per group). (D) Schema showing how excess immature HSPCs may yield normal marrow cellularity and blood cell counts due to apoptosis of surplus hematopoietic precursors. (E) Confocal immunofluorescence imaging of cleaved Caspase 3 (Casp3; green, Cy3) and Cdh5 (magenta, Alexa Fluor 647) in BM sections of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 3 per group). Original magnification ×20. (F) Representative flow cytometry data of cleaved Caspase 3/7 activity is shown for TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice and for the FMO− control. (G) Quantification of apoptotic cells was assessed using the Caspase 3/7 reporter (n = 3 per group). (H) Representative flow cytometry data showing annexin V staining in marrow from TGFβ1FL/FL (top row) and TGFβ1ΔMk/ΔMk (bottom row) mice. (I) Quantification of the number of annexin V+ apoptotic cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 9 per group). (J) Representative data for spleen, as shown for marrow in panel H. (K) Quantification of the number of annexin V+ apoptotic cells in the spleen of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 6 per group). (L) Representative flow cytometry data showing lineage markers (Lin = CD3, B220, CD11b, Gr1, or Ter119) plotted against annexin V in marrow from TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice. (M) Quantification of the number of annexin V+ apoptotic Lin+ and Lin− cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 8). (N) Gating and representative flow cytometry data showing annexin V staining in LKS− and LKS+ marrow populations for TGFβ1FL/FL (top row) and TGFβ1ΔMk/ΔMk (bottom row) mice. (O) Quantification of the number of annexin V+ apoptotic LKS− and LKS+ cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 3 per group). All the quantified data are shown as mean plus or minus standard deviation (SD) (*P < .05, **P < .01, ***P < .001, or if not shown, the comparison was not significant). FSA, forward scatter area; TNC, total nucleated cell; VECAD, vascular endothelial cadherin.

Conditional deletion of TGFβ1 in Mks increases apoptosis. (A) White blood cell (WBC), RBC, and platelet (PLT) counts are shown for TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 16 per group). (B) Marrow (n = 12 per group) and spleen (n = 5 per group) cellularity are shown for TGFβ1FL/FL (gray) and TGFβ1ΔMk/ΔMk mice (black). (C) The number of Lin− (those not expressing mature lineage markers CD3, B220, CD11b, Gr1, or Ter119) marrow cells per leg (femur and tibia) is shown (n = 13 per group). (D) Schema showing how excess immature HSPCs may yield normal marrow cellularity and blood cell counts due to apoptosis of surplus hematopoietic precursors. (E) Confocal immunofluorescence imaging of cleaved Caspase 3 (Casp3; green, Cy3) and Cdh5 (magenta, Alexa Fluor 647) in BM sections of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 3 per group). Original magnification ×20. (F) Representative flow cytometry data of cleaved Caspase 3/7 activity is shown for TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice and for the FMO− control. (G) Quantification of apoptotic cells was assessed using the Caspase 3/7 reporter (n = 3 per group). (H) Representative flow cytometry data showing annexin V staining in marrow from TGFβ1FL/FL (top row) and TGFβ1ΔMk/ΔMk (bottom row) mice. (I) Quantification of the number of annexin V+ apoptotic cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 9 per group). (J) Representative data for spleen, as shown for marrow in panel H. (K) Quantification of the number of annexin V+ apoptotic cells in the spleen of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 6 per group). (L) Representative flow cytometry data showing lineage markers (Lin = CD3, B220, CD11b, Gr1, or Ter119) plotted against annexin V in marrow from TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice. (M) Quantification of the number of annexin V+ apoptotic Lin+ and Lin− cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 8). (N) Gating and representative flow cytometry data showing annexin V staining in LKS− and LKS+ marrow populations for TGFβ1FL/FL (top row) and TGFβ1ΔMk/ΔMk (bottom row) mice. (O) Quantification of the number of annexin V+ apoptotic LKS− and LKS+ cells in the marrow of TGFβ1FL/FL and TGFβ1ΔMk/ΔMk mice (n = 3 per group). All the quantified data are shown as mean plus or minus standard deviation (SD) (*P < .05, **P < .01, ***P < .001, or if not shown, the comparison was not significant). FSA, forward scatter area; TNC, total nucleated cell; VECAD, vascular endothelial cadherin.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal