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Fig. 1 | Biology of Sex Differences

Fig. 1

From: Sex differences in the percentage of IRF5 positive B cells are associated with higher production of TNF-α in women in response to TLR9 in humans

Fig. 1

Sex differences in the percentages of IRF5 positive B cells. A Flow cytometric plots showing the applied gating strategy for isolated B cells (CD3–CD20 + cells). FACS plots of a representative donor are shown (out of 22 experiments). B cells were purified with Human B Cell Enrichment Cocktail. The percentages of IRF5 positive B cells were analyzed ex vivo by flow cytometry. The first gate was set on physical parameters, then on SSC-W versus SSC-A and FSC-W versus FSC-A to eliminate doublets, then on CD3 negative events to exclude remaining CD3 positive cells. IRF5 levels (in blue) were determined by the use of an isotype control (in red) in B cells from women or age-matched men. B To confirm antibody specificity, we performed surface staining to detect IRF5 on the cell surface of CD20 + B cells. IRF5 staining was performed by surface staining (orange histogram) and after permeabilized in 0.01% Triton-X-100 for intracellular staining (blue histogram) or isotype control (red histogram). Surface staining of IRF5 served as a negative control as IRF5 is located in the cytoplasm and intranuclearly [25]. No IRF5 positive B cells were detected after surface staining alone. C Percentages of IRF5 positive B cells. Women (pink dots) showed significantly higher percentage of IRF5 positive B cells than men (blue dots) in unstimulated conditions (p = 0.01; two-tailed t test; women n = 11, men n = 11)

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