Figure 5
Identification of interstitial and glomerular renal macrophages using reporter lines and antibodies. (A–D) Dual fluorescence for GFP and macrophage antibodies in kidney sections from CSF1R-EGFP mice. In the tubulointerstititial space ~ 50% of CSF1R+ cells (white arrows) are identified by Mac2 (A) and Mac3 antibodies (B). Moreover, virtually all CSF1R+ cells are stained for F4/80 (CI:A3-1 clone) (C, arrows). F4/80 BM8 clone also identifies renal macrophages; in contrast no staining was obtained with ab 111,101 (Supplemental Fig. I). CSF1R+ renal macrophages were not labeled with the anti-lysozyme antibody clone EPR2994(2) (D). A subset of tubular epithelial cells shows intense staining for Mac 2 (A, yellow arrows) and lysozyme M (yellow arrows, D). The antibody to Mac3 stains the brush-border of the epithelial cells (B, yellow arrows). (E–H) Dual fluorescence for GFP and macrophage antibodies in kidney sections from CX3CR1GFP mice. A small fraction of CX3CR1+ cells express Mac2+ (E) or Mac3+ (F). Virtually all CX3CR1+ cells stain for F4/80 (clone CI:A3-1) (G, arrows). CX3CR1+ cells do not stain with the lysozyme antibody, which labels intensely the tubular epithelium (H). I-P: Images illustrating staining of glomerular macrophages. In glomeruli, few CSF1R+ (I-L) and CX3CR1+ cells (M–P) are noted. The majority of these cells are labeled with Mac2 (I, M), and F4/80 CI:A3-1 clone (K, O—arrows), but not with Mac3 (J, N), and anti-Lysozyme antibodies (L, P). Scale bar: 20 μm.
