Figure 7.
BM-SECs are a site of Fgf23 upregulation in genetically induced IDA, in phlebotomy-induced anemia, and after the direct administration of EPO. (A) Immunohistochemical staining with an anti-GFP antibody in the BM of formalin-fixed, decalcified femur sections of selected Tmprss6-Fgf23 genotype combinations (6-month-old male mice). Original magnification ×40. Brown staining highlights anti-GFP immunoreactivity; blue staining reflects hematoxylin counterstain. (B-E) Circulating parameters in Fgf23+/eGFP and Fgf23+/+ mice evaluated at the 18-hour time point after a large-volume phlebotomy, compared with nonphlebotomized, genotype-matched controls, validating the expected effects of phlebotomy on (B) Hgb, (C) red blood cell (RBC) count, (D) serum EPO, and (E) plasma cFGF23. Fgf23+/eGFP and Fgf23+/+ female mice (8-weeks-old; 2-4 per group) were either not previously phlebotomized (NP) or underwent a single 500 μL phlebotomy with saline volume replacement 18 hours earlier ("Phleb"). For all graphs, data represent mean ± standard deviation. ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001 using a two-way ANOVA with Tukey post hoc test. (F) Immunohistochemical staining with anti-GFP antibody in the BM of formalin-fixed, decalcified femur sections harvested 18 hours after a 500 μL phlebotomy (Fgf23+/+ or Fgf23+/eGFP mice) or harvested from nonphlebotomized Fgf23+/eGFP control mice. Original magnification ×40. (G) Immunohistochemical staining with anti-GFP antibody in the BM of formalin-fixed decalcified femur sections harvested 6 hours after injection of EPO (Fgf23+/+ or Fgf23+/eGFP mice) or saline vehicle (Fgf23+/eGFP control mice). Original magnification ×40.

BM-SECs are a site of Fgf23 upregulation in genetically induced IDA, in phlebotomy-induced anemia, and after the direct administration of EPO. (A) Immunohistochemical staining with an anti-GFP antibody in the BM of formalin-fixed, decalcified femur sections of selected Tmprss6-Fgf23 genotype combinations (6-month-old male mice). Original magnification ×40. Brown staining highlights anti-GFP immunoreactivity; blue staining reflects hematoxylin counterstain. (B-E) Circulating parameters in Fgf23+/eGFP and Fgf23+/+ mice evaluated at the 18-hour time point after a large-volume phlebotomy, compared with nonphlebotomized, genotype-matched controls, validating the expected effects of phlebotomy on (B) Hgb, (C) red blood cell (RBC) count, (D) serum EPO, and (E) plasma cFGF23. Fgf23+/eGFP and Fgf23+/+ female mice (8-weeks-old; 2-4 per group) were either not previously phlebotomized (NP) or underwent a single 500 μL phlebotomy with saline volume replacement 18 hours earlier ("Phleb"). For all graphs, data represent mean ± standard deviation. ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001 using a two-way ANOVA with Tukey post hoc test. (F) Immunohistochemical staining with anti-GFP antibody in the BM of formalin-fixed, decalcified femur sections harvested 18 hours after a 500 μL phlebotomy (Fgf23+/+ or Fgf23+/eGFP mice) or harvested from nonphlebotomized Fgf23+/eGFP control mice. Original magnification ×40. (G) Immunohistochemical staining with anti-GFP antibody in the BM of formalin-fixed decalcified femur sections harvested 6 hours after injection of EPO (Fgf23+/+ or Fgf23+/eGFP mice) or saline vehicle (Fgf23+/eGFP control mice). Original magnification ×40.

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