pe cyanine7 pe cy7 conjugated cd11b  (Thermo Fisher)

Journal: eLife

Article Title: Hemozoin produced by mammals confers heme tolerance

doi: 10.7554/eLife.49503

Figure Lengend Snippet: Quantifications of total Ter-119 + cells in the ( B ) bone marrow and ( E ) spleen. The %single cells* on the y-axis denote single cells that are negative for CD4/8/41, B220 and Gr-1. ( C ) Quantification of subpopulations of Ter-119 + cells represented as a percentage of total Ter-119 + cells in the bone marrow (n = 7–12). ( F ) Quantification of populations II and III of Ter-119 + cells represented as a percentage of total Ter-119 + cells in the spleen. Gating strategy ( G, I ) and quantification ( H, J ) of ( G, H ) splenic F4/80 hi Treml4 + red pulp macrophages (RPMs) and ( I, J ) F4/80 hi and F4/80 lo -CD11b hi splenic monocytes. At least 100,000 single cells were analyzed per sample. Each dot represents one mouse. **p<0.05; **p<0.01.

Article Snippet: For non-erythroid cell analysis, splenic cells were treated with RBC lysis buffer and stained with the following antibodies: phycoerythrin (PE)-conjugated Treml4 (Biolegend, San Diego, CA, cat. number 143304), APC-conjugated F4/80 (eBioscience, cat. number 17-4801-80), and PE-Cyanine7 (PE-Cy7)-conjugated CD11b (eBioscience, cat. number 25-0112-81).

Techniques:

Journal: eLife

Article Title: Hemozoin produced by mammals confers heme tolerance

doi: 10.7554/eLife.49503

Figure Lengend Snippet: ( A ) Kaplan-Meier survival curve of WT and KO mice placed on a low-iron (2ppm) diet (n = 15–17, both males and females). ( B–C ) Hematocrits of WT and KO mice placed on a standard or low-iron (2ppm) diet. Mice were placed on respective diets supplemented with deionized water starting at 21 days of age (week 0) (n = 9–15 for 5 week data set; n = 7–11 for 20 week data set). Quantification of tissue iron ( D ) and heme ( E ) by ICP-MS and UPLC, respectively, in tissues of mice fed a low-iron (2ppm) diet (n = 6–17). ( F ) %Splenomegaly of WT and KO mice calculated by the percentage of increase in average wet weight of spleens between mice on low-iron versus standard iron diets (n = 9–15); ( G ) Ratio of 2ppm splenic Ter-119 + population II+III cells to that of standard diet mice (n = 8–12); ( H ) Quantification of total Ter-119 + cells in the spleen. The %single cells* on the y-axis denote single cells that are negative for CD4/8/41, B220 and Gr-1. Each point represents one mouse. ( I ) Quantification of subpopulations of Ter-119 + cells represented as a percentage of total Ter-119 + cells in the bone marrow (n = 7–8); ( J ) Quantifications of splenic RPMs in mice on a standard or low-iron (2ppm) diet, represented as a percentage of single cells analyzed (n = 9–14). ( K ) Quantification of the ratio of F4/80 hi to F4/80 lo CD11b hi splenic monocytes from mice on a standard or low-iron (2ppm) diet (n = 9–15). At least 100,000 single cells were analyzed per sample. ( L ) Gene expression heat map of 90 iron metabolism genes in spleens from mice on standard or low-iron (2ppm) diet. Pearson correlation was used for comparison; average linkage (n = 9 per group, per genotype). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Article Snippet: For non-erythroid cell analysis, splenic cells were treated with RBC lysis buffer and stained with the following antibodies: phycoerythrin (PE)-conjugated Treml4 (Biolegend, San Diego, CA, cat. number 143304), APC-conjugated F4/80 (eBioscience, cat. number 17-4801-80), and PE-Cyanine7 (PE-Cy7)-conjugated CD11b (eBioscience, cat. number 25-0112-81).

Techniques: Expressing