mouse anti rat cd68 conjugated with pe  (Novus Biologicals)

Journal: Frontiers in Immunology

Article Title: Site-Specific Reprogramming of Macrophage Responsiveness to Bacterial Lipopolysaccharide in Obesity

doi: 10.3389/fimmu.2019.01496

Figure Lengend Snippet: Macrophage abundance and cytokine secretion in AT-SVF cultures. (A,B) Representative flow-cytometry density plots of staining for CD45 (a leukocyte marker) and CD68 (a rat macrophage marker) in AT-SVF cells from LFD-fed rats (A) or HFD-fed rats (B) . Stained cells are shown in red; unstained cells are shown in gray. Group results— n = 5 culture replicates in (A) ; n = 12 in (B) —are shown as 95% confidence intervals within each quadrant. (C) Representative histograms showing that LPS-stimulated cells that are CD45 + /CD68 + (macrophages) stain more strongly for TNF-α than the other cellular phenotypes present in the AT-SVF culture. For staining intensities of all samples, see . (D–F) Effects of diet and ex-vivo stimulation with LPS on the secretion of TNF-α (D) , IL-1β (E) and IL-10 (F) by AT-SVF cells. To aid comparison of this macrophage subpopulation with the others, the scales in these panels are identical to those of the corresponding panels in , . Data in (D–F) are expressed as means ± SEM. Sample size (number of culture replicates) was 4 in the LFD group not stimulated with LPS (saline), 6 in the HFD group not stimulated with LPS (saline), 5 in the LFD group stimulated with LPS, and 7 in the HFD group stimulated with LPS. (G) TNF-α/IL-10 and IL-1β/IL-10 ratios (means ± SEM) in the LPS-stimulated AT-SVF cultures from the LFD vs. HFD groups. Statistical marks: a significant effect of LPS (compared to saline), b significant effect of diet.

Article Snippet: Stainings were achieved by 30-min incubation with the following monoclonal antibodies at 4°C and in the dark: mouse anti-rat CD45 conjugated with APC-Cy7 (BD 561586; 1:100); mouse anti-rat CD68 conjugated with PE (Novus Biologicals NB600-985PE; 1:100); and hamster anti-rat/mouse TNF-α conjugated with APC (BioLegend 506107; 1:25).

Techniques: Flow Cytometry, Staining, Marker, Ex Vivo, Comparison, Saline

Journal: Frontiers in Immunology

Article Title: Site-Specific Reprogramming of Macrophage Responsiveness to Bacterial Lipopolysaccharide in Obesity

doi: 10.3389/fimmu.2019.01496

Figure Lengend Snippet: Macrophage abundance and cytokine secretion in alveolar cultures. (A,B) Representative flow-cytometry density plots of staining for CD45 (a leukocyte marker) and CD68 (a rat macrophage marker) in alveolar cells from LFD-fed rats (A) or HFD-fed rats (B) . Stained cells are shown in red; unstained cells are shown in gray. Group results— n = 5 culture replicates in (A) ; n = 8 in (B) —are shown as 95% confidence intervals within each quadrant. (C) Representative histograms showing that LPS-stimulated cells that are CD45 + /CD68 + (macrophages) stain strongly for TNF-α. Because CD45 + /CD68 − were very rare in the alveolar culture, their histograms could not be reliably determined and, consequently, this subpopulation is not shown in (C) of this particular figure. For staining intensities of all samples, see . (D–F) Effects of diet and ex-vivo stimulation with LPS on the secretion of TNF-α (D) , IL-1β (E) , and IL-10 (F) by alveolar macrophages. To aid comparison of this macrophage subpopulation with the others, the scales in these panels are identical to those of the corresponding panels in , . Data in (D–F) are expressed as means ± SEM. Sample size (number of culture replicates) was 6 in the LFD group not stimulated with LPS (saline), 7 in the HFD group not stimulated with LPS (saline), 18 in the LFD group stimulated with LPS, and 16 in the HFD group stimulated with LPS. (G) TNF-α/IL-10 and IL-1β/IL-10 ratios (means ± SEM) in the LPS-stimulated alveolar macrophages from the LFD vs. HFD groups. Statistical mark: a significant effect of LPS (compared to saline).

Article Snippet: Stainings were achieved by 30-min incubation with the following monoclonal antibodies at 4°C and in the dark: mouse anti-rat CD45 conjugated with APC-Cy7 (BD 561586; 1:100); mouse anti-rat CD68 conjugated with PE (Novus Biologicals NB600-985PE; 1:100); and hamster anti-rat/mouse TNF-α conjugated with APC (BioLegend 506107; 1:25).

Techniques: Flow Cytometry, Staining, Marker, Ex Vivo, Comparison, Saline

Journal: Frontiers in Immunology

Article Title: Site-Specific Reprogramming of Macrophage Responsiveness to Bacterial Lipopolysaccharide in Obesity

doi: 10.3389/fimmu.2019.01496

Figure Lengend Snippet: Macrophage abundance and cytokine secretion in peritoneal cultures. (A,B) Representative flow-cytometry density plots of staining for CD45 (a leukocyte marker) and CD68 (a rat macrophage marker) in peritoneal cells from LFD-fed rats (A) or HFD-fed rats (B) . Stained cells are shown in red; unstained cells are shown in gray. Group results— n = 5 culture replicates in (A) ; n = 8 in (B) —are shown as 95% confidence intervals within each quadrant. (C) Representative histograms showing that LPS-stimulated cells that are CD45 + /CD68 + (macrophages) stain more strongly for TNF-α than the other cellular phenotypes present in the peritoneal culture. For staining intensities of all samples, see . (D–F) Effects of diet and ex-vivo stimulation with LPS on the secretion of TNF-α (D) , IL-1β (E) , and IL-10 (F) by peritoneal macrophages. To aid comparison of this macrophage subpopulation with the others, the scales in these panels are identical to those of the corresponding panels in , . Data are expressed as means ± SEM. Sample size (number of culture replicates) was 12 in the LFD group not stimulated with LPS (saline), 7 in the HFD group not stimulated with LPS (saline), 16 in the LFD group stimulated with LPS, and 13 in the HFD group stimulated with LPS. (G) TNF-α/IL-10 and IL-1β/IL-10 ratios (means ± SEM) in the LPS-stimulated peritoneal macrophages from the LFD vs. HFD groups. Statistical marks: a significant effect of LPS (compared to saline), b significant effect of diet.

Article Snippet: Stainings were achieved by 30-min incubation with the following monoclonal antibodies at 4°C and in the dark: mouse anti-rat CD45 conjugated with APC-Cy7 (BD 561586; 1:100); mouse anti-rat CD68 conjugated with PE (Novus Biologicals NB600-985PE; 1:100); and hamster anti-rat/mouse TNF-α conjugated with APC (BioLegend 506107; 1:25).

Techniques: Flow Cytometry, Staining, Marker, Ex Vivo, Comparison, Saline