Journal: The American Journal of Pathology

Article Title: Colony-Stimulating Factor-1 Promotes Kidney Growth and Repair via Alteration of Macrophage Responses

doi: 10.1016/j.ajpath.2011.05.037

Figure Lengend Snippet: Macrophage infiltration is increased in kidneys at P5 after treatment with CSF-1. A–D: Flow cytometric analysis of whole kidneys indicates CD45+CD11b+CD11c−/F480+ renal macrophages are increased in CSF-1-treated mice at P5. Representative histograms show an increase in CD45+CD11b+CD11c− cells expressing the macrophage marker F4/80 with CSF-1 treatment (B) compared with littermate vehicle-injected controls (A). C: Positive staining was confirmed with appropriate isotype controls. E—G: Representative images with the use of fluorescent microscopy show csf1r-EGFP macrophage infiltration (arrows) in kidneys at P5 after treatment with either CSF-1 or vehicle control at P1, P2, and P3 (n = 4 per group). Nuclei are stained with DAPI (blue). Low magnification (×100) shows that kidneys treated with CSF-1 (F) had increased numbers of macrophages in the renal medulla and cortex, compared with vehicle controls (E). G:Csf1r-EGFP macrophages show a spindle-shaped structure, wrapping around newly developed structures. H: Estimation of the number of renal csf1r-EGFP macrophages showed a 42% (P = 0.004) increase in macrophages within the cortex and a 157% (P = 0.003) increase within the medulla after treatment with CSF-1, compared with vehicles (n = 3 to 4 per group). Data were analyzed with a Student's t-test (unpaired, 2-tailed); *P < 0.05, **P < 0.01. Data are means ± SEMs. Veh, vehicle.

Article Snippet: Cells were adjusted to 3 × 10 6 cells/test in 96-well trays and stained with the following mAb cocktail for 15 minutes at 4°C: CD45-APC Cy7 (clone 30-F11), CD11c-Pacific Blue, (clone N418), mannose receptor (MR) CD206-Alexa Fluor 488 (clone MR5D3; BioLegend, San Diego, CA); CD11b-PE Cy7 (clone M1/70) MHCII/1-A/1-E-PE (Clone M5/114.15.2; BD Biosciences); and F4/80-APC (clone BM8; eBioscence, San Diego, CA).

Techniques: Expressing, Marker, Injection, Staining, Microscopy

Journal: The American Journal of Pathology

Article Title: Colony-Stimulating Factor-1 Promotes Kidney Growth and Repair via Alteration of Macrophage Responses

doi: 10.1016/j.ajpath.2011.05.037

Figure Lengend Snippet: CD45+CD11b+CD11c− renal macrophages in CSF-1-treated mice display an altered phenotype and cell number compared with vehicle-treated mice at 5 and 7 days after IR injury. Flow cytometric analysis was performed on sham-operated kidneys and kidneys 5 and 7 days after IR treated with vehicle or CSF-1. A: Flow cytometric quantification of leukocytes stained for F480, MR, and MHC class II on CD45+CD11b+CD11c− renal macrophages. B: Representative histograms displaying the shift in mean fluorescent intensity of MHC class II+ cells gated on CD45+CD11b+CD11c−/F480+ macrophages. C: Representative contour plots of F480 and MR co-expression on CD45+CD11b+CD11c− macrophages. Positive staining was confirmed with appropriate isotype controls. Percentages of cells in each quadrant represent mean ± SEM. Statistical analysis was performed with a one-way analysis of variance with an accompanying Tukey's post hoc test for multiple comparisons; *P < 0.05, **P < 0.01, and ***P < 0.001. Data are means ± SEMs (n = 5 per group). MFI, mean fluorescent intensity; Veh, vehicle.

Article Snippet: Cells were adjusted to 3 × 10 6 cells/test in 96-well trays and stained with the following mAb cocktail for 15 minutes at 4°C: CD45-APC Cy7 (clone 30-F11), CD11c-Pacific Blue, (clone N418), mannose receptor (MR) CD206-Alexa Fluor 488 (clone MR5D3; BioLegend, San Diego, CA); CD11b-PE Cy7 (clone M1/70) MHCII/1-A/1-E-PE (Clone M5/114.15.2; BD Biosciences); and F4/80-APC (clone BM8; eBioscence, San Diego, CA).

Techniques: Staining, Expressing

Journal: The American Journal of Pathology

Article Title: Colony-Stimulating Factor-1 Promotes Kidney Growth and Repair via Alteration of Macrophage Responses

doi: 10.1016/j.ajpath.2011.05.037

Figure Lengend Snippet: qPCR analysis of pro-inflammatory and anti-inflammatory gene expression in sorted CD45+CD11b+CD11c− macrophages from sham-operated kidneys and kidneys 5 and 7 days after IR treated with CSF-1 or vehicle injections. Pro-inflammatory (INOS, TNF-α, CCL2, and Cxcl2) (A) and anti-inflammatory (CCL17, arginase, MSR2, and MR) (B) gene expression in response to CSF-1 treatment. Sham-operated kidneys were used as the baseline control. Statistical analysis was performed with a one-way analysis of variance with an accompanying Tukey's post hoc test for multiple comparisons; *P < 0.05, **P < 0.01, and ***P < 0.001. Data are means ± SEMs (n = 5 per group). RQ, relative quantification; Veh, vehicle.

Article Snippet: Cells were adjusted to 3 × 10 6 cells/test in 96-well trays and stained with the following mAb cocktail for 15 minutes at 4°C: CD45-APC Cy7 (clone 30-F11), CD11c-Pacific Blue, (clone N418), mannose receptor (MR) CD206-Alexa Fluor 488 (clone MR5D3; BioLegend, San Diego, CA); CD11b-PE Cy7 (clone M1/70) MHCII/1-A/1-E-PE (Clone M5/114.15.2; BD Biosciences); and F4/80-APC (clone BM8; eBioscence, San Diego, CA).

Techniques: Expressing