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Application Data (Published customer image Infiltration of GFP+ BM-cells in infarct and peri-infarct regions. (A-B) Dot plots of viable macrophages/granulocytes (CD11b+CD45high, top right quadrants) and microglia (CD11b+CD45dim, bottom right quadrants) in cortex from BM-chimeric unmanipulated mice and mice exposed to pMCAO. (C) Bar graph showing mean numbers of CD11b+CD45dim microglia and CD11b+CD45high macrophages/granulocytes in BM-chimeric mice 24 hours after pMCAO, subdivided based on expression of GFP (n = 5). Approximately 92% of of the CD45high population were GFP+. (D) Estimation and comparison of mean numbers of CD11b+CD45dim microglia in non-chimeric (n = 10) versus BM-chimeric mice (n = 5) 24 hours after of pMCAO shows significantly fewer CD11b+CD45dim microglial cells in irradiated mice. (E) Overview, showing distribution of infiltrating GFP+ BM-derived cells into infarct (IF) and peri-infarct (P-IF) regions 24 hours after pMCAO. (E-G) By 24 hours, GFP+ single cells (F) and vessel-associated aggregates of GFP+ cells (arrows in G) were observed in infarct and peri-infarct regions. Some of the vessel-associated cells were round, leukocyte-like cells (arrows) while others were elongated cells lining the vasculature (arrow heads in G and in insert). (H) Bar graph showing mean numbers of single GFP+ cells and vessel-associated aggregates of GFP+ cells in ipsi- and contralateral cortex 24 hours after surgery (n = 10). (I-P) Immunohistochemical staining of CD45.1 (I, K), CD45.2 (J, L), IgG2a (M, O) and CD45 (N, P) in ischemic tissue in BM-chimeric (I, J, M, N) and non-chimeric mice (K, L, O, P) 24 hours after pMCAO. N.D, none detected. Scale bars: 200 um (A), 10 um (B, C). 50 um (I-P) *P < 0.05, **P < 0.01, and ***P < 0.001.From: Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B. Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice. J Neuroinflammation. 2008 Oct 23;5:46.)

Application Data (Staining of mouse bone marrow with Rat anti Mouse CD11b)

Application Data (Immunoperoxidase staining o mouse lymph node cryosection with Rat anti Mouse CD11b antibody, clone 5C6 followed by horseradish peroxidase conjugated Goat anti Rat IgG as a detection reagent. Low power)

Application Data (Published customer image: Activation of adult primary microglial cells in wild-type and IL-1 KO mice. (A) Primary microglial cells were obtained from young adult wild-type mice. The cells stain with the microglial marker CD11b, but not with the neuronal and astroglial markers, NeuN and GFAP, respectively. A few cells are stained with the oligodendroglial cell marker, MBP. NC (inset) is the primary antibody-free negative control. The microglial cells (n = 3 each group) were stimulated for 24 hours in the presence of the vehicle alone, or supplemented with IFN? or IL-4 in the presence or absence of IL-1beta. Total NO (NOx; B), TNFa (C), arginase specific activity (Arg-1 spe. act.; D) and IGF-1 (E) were determined from the media or cell suspensions. (B) NOx levels increase upon exposure of the cells to IL-1beta and in a synergistic manner upon co-treatment of cells with IL-1beta and IFN?, but not when the cotreatment is with IL-4. (C) TNFa levels increase upon exposure of the cells to IFN?, and further upon co-treatment with IL-1beta. Surprisingly, the co-treatment of the cells with IL-4 and IL-1beta induced the highest TNFa level among the experimental treatments used. (D) Arg1-specific activity increased significantly upon exposure to IL-4 and further increased when IL-4 and IL-1beta were employed together. (E) IGF-1 levels decreased with exposure of the cells to IFN? and increased in response to IL-4. The response was partially inhibited by cotreatment of the cells with IL-1beta. Data are expressed as mean +/- SD (n = 3). *: P < 0.05, **: P < 0.01, ***: P < 0.001 compared with the vehicle-treated group in each genotype (one-way ANOVA followed by Dunnett post-hoc test). ANOVA, analysis of variance; IGF-1, insulin-like growth factor.From: Sato A, Ohtaki H, Tsumuraya T, Song D, Ohara K, Asano M, Iwakura Y, Atsumi T, Shioda S. Interleukin-1 participates in the classical and alternative activation of microglia/macrophages after spinal cord injury. J Neuroinflammation. 2012 Apr 7;9:65. doi: 10.1186/1742-2094-9-65.)

Application Data (Staining of mouse peritoneal macrophages with Rat anti Mouse CD11b:FITC)

Application Data (Immunoperoxidase staining of mouse lymph node cryosection with Rat anti Mouse CD11b antibody, clone 5C6 followed by horseradish peroxidase conjugated Goat anti Rat IgG as a detection reagent. high power)

Application Data (Published customer image BM-derived GFP+ single cells and vessel-associated cells express CD11b. Fluorescence microscopy for GFP combined with immunofluorescence detection of (A) CD11b, (B) vWF and (C) CD31, 24 hours after pMCAO. (A) Fluorescence detection of GFP and CD11b showed that most GFP+ cells co-expressed CD11b (yellow cells, indicated by arrows), and intermingled with CD11b+ host cells. Note also that a few GFP+ cells did not co-express CD11b (arrow head). Insert shows high magnification of GFP+ cells, some of which co-express CD11b, aggregated around a vessel. (B, C) Fluorescence detection of GFP and the endothelial cell markers vWF (B) and CD31 (C). Inserts show higher magnification of sections of the same vessels. Although there are indications that single vWF+ cells co-express GFP (arrows in B), this could not be reproduced using staining for CD31, and the majority of vWF+ and CD31+ cells showed no co-expression of GFP. Instead, GFP remained confined to round and elongated cells located in the juxtavascular space (insert in C). CD11b+ cells were visualized using Alexa Fluor 568-conjugated goat anti-rat IgG, vWF+ and CD31+ cells using Alexa Fluor 546-conjugated goat anti-rabbit IgG and Alexa Fluor 594-conjugated goat anti-rat IgG, respectively. Scale bars: 20 um (A-C)..From: Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B. Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice. J Neuroinflammation. 2008 Oct 23;5:46.)

Application Data (Immunofluorescence staining of mouse lymph node cryosection with Rat anti Mouse CD11b, clone 5C6 , green in A and Rat anti Mouse CD8, clone YTS105.18 , red in B. C is the merged image with nuclei counterstained blue using DAPI. High power)