Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2009 Dec 15.

Published in final edited form as: J Immunol. 2008 Dec 15;181(12):8670–8676. doi: 10.4049/jimmunol.181.12.8670

A–E) Immunohistochemical analysis. A) Macrophages. anti-F4/80 staining of wild type C57BL/6 (i) and CCR1-deficient (ii) mouse kidney, and control IgG staining of wild C57BL/6 mouse kidney (iii). B) Neutrophils. naphthol AS-D chloroacetate esterase staining of wild type C57BL/6 (i) and CCR1-deficient (ii) mouse kidney. Original magnification of all images is 320X. Scale bar indicates 50µm. C–E, Quantitation of density of indicated leukocyte subsets in tissue sections at 320X magnification under the conditions shown. sham, sham-operated wild type mice tested 24 hours after surgery. Values are mean ± SEM and are pooled from 3 independent experiments with 3–5 animals for each condition in each experiment. * P<0.05 comparing CCR1-deficient mice versus wild type mice at the same time point. F–I, FACS analysis. Representative FACS data, in which anti-CD45 (Fi, ii) or isotype control (Fiii, iv) was used to stain cells isolated from whole left injured kidney (Fi, iii) or right control kidney (Fii, iv) 7 days post-IR. (Fv-viii) Representative FACS data for CD45+ cells purified using immunobeads and stained with anti-F4/80 (v), anti-Gr-1 (vi), anti-CD3 (vii), or isotype control (viii). G, Summary data of FACS analysis. % of total was calculated as (% of CD45+) X (% of F4/80+, Gr-1+ or CD3+ cells) in each animal. n=8 for CCR1-deficient mice, n=14 for C57BL/6 control mice. H, Representative FACS data 7 days after reperfusion, in which PE conjugated anti-CCR1 (Hii-iv) or control IgG (Hi) and FITC conjugated anti-F4/80(Hiii), anti-GR1(Hiv) or isotype control (Hii) were used to stain CD45-positive cells isolated from whole left injured kidney by magnetic cell sorting methods. I, Summary data of FACS analysis. % of F4/80- or Gr-1-positive cells in CCR1-positive cells. n=5 in each group.

A–E) Immunohistochemical analysis. A) Macrophages. anti-F4/80 staining of wild type C57BL/6 (i) and CCR1-deficient (ii) mouse kidney, and control IgG staining of wild C57BL/6 mouse kidney (iii). B) Neutrophils. naphthol AS-D chloroacetate esterase staining of wild type C57BL/6 (i) and CCR1-deficient (ii) mouse kidney. Original magnification of all images is 320X. Scale bar indicates 50µm. C–E, Quantitation of density of indicated leukocyte subsets in tissue sections at 320X magnification under the conditions shown. sham, sham-operated wild type mice tested 24 hours after surgery. Values are mean ± SEM and are pooled from 3 independent experiments with 3–5 animals for each condition in each experiment. * P<0.05 comparing CCR1-deficient mice versus wild type mice at the same time point. F–I, FACS analysis. Representative FACS data, in which anti-CD45 (Fi, ii) or isotype control (Fiii, iv) was used to stain cells isolated from whole left injured kidney (Fi, iii) or right control kidney (Fii, iv) 7 days post-IR. (Fv-viii) Representative FACS data for CD45+ cells purified using immunobeads and stained with anti-F4/80 (v), anti-Gr-1 (vi), anti-CD3 (vii), or isotype control (viii). G, Summary data of FACS analysis. % of total was calculated as (% of CD45+) X (% of F4/80+, Gr-1+ or CD3+ cells) in each animal. n=8 for CCR1-deficient mice, n=14 for C57BL/6 control mice. H, Representative FACS data 7 days after reperfusion, in which PE conjugated anti-CCR1 (Hii-iv) or control IgG (Hi) and FITC conjugated anti-F4/80(Hiii), anti-GR1(Hiv) or isotype control (Hii) were used to stain CD45-positive cells isolated from whole left injured kidney by magnetic cell sorting methods. I, Summary data of FACS analysis. % of F4/80- or Gr-1-positive cells in CCR1-positive cells. n=5 in each group.