Figure 3

KRM repopulate from donor bone marrow (BM) cells but do not sustain in stenotic kidneys resulting in loss of peritubular capillaries. (A) Experimental Schema for BM transplantation (BMT) studies. (B–D) Reconstitution by donor-derived (CD45.1) BM in BMT + Sham, BMT + RAS stenotic and BMT + RAS contralateral kidneys (Top Row); Native (black tinted) and donor-derived macrophages (pink tinted) (Middle Row); Donor-derived KRM population decreased in BMT + RAS (Bottom Row). (E) Immunostaining of PLVAP and CD31 showing peri-tubular endothelial cells in representative images of BMT + Sham and BMT + RAS kidney sections. Images acquired on Zeiss confocal at 40X and stitched together to show a larger area. Note CD31 (red) stains peri-glomerular cells while PLVAP (green) is specific to peri-tubular endothelial cells. (F) Quantifying PLVAP⁺CD31⁺ cells showing significant reduction in BMT + RAS Vs BMT + Sham. (G) Experimental Schema for administration of liposomal clodronate at low-doses. (H) Comparing percent of all three macrophages in the stenotic kidneys of Sham, RAS, RAS + Vehicle and RAS + Clodronate mice. Note significant reduction in KRM in RAS + Clodronate group. (I) Immunostaining of PLVAP and CD31 showing peri-tubular endothelial cells in representative images of RAS + Vehicle and RAS + Clodronate kidney sections. (J) Quantifying PLVAP⁺CD31⁺ cells. Significant loss of peritubular endothelial cells seen after administration of clodronate. n = 6 mice/group; *P ≤ 0.01 vs Sham; ^§P < 0.05 vs BMT + Sham , RAS + Vehicle; ^¥P < 0.01 vs RAS. Mouse images adopted from Openclipart.org https://openclipart.org/detail/174870/mouse and https://openclipart.org/detail/28929/kidneyreins.