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. 2017 Aug 25;8:1044. doi: 10.3389/fimmu.2017.01044

Figure 4.

Figure 4

Proliferative responses of B cell subsets after stimulation with toll-like receptor (TLR) ligands (A) Gating strategy defining CD21+IgM+, CD21+IgM+, and IgG+ B cells, exemplified for unstimulated and Pam3Cys-SK4 stimulated peripheral blood mononuclear cells (PBMCs) after 5 days of culture. (B) CellTrace Violet staining of the three B-cell subsets defined in (A) using multicolor flow cytometry. PBMCs were cultured for 5d. A representative animal out of three is shown. (C) Percentage of proliferating B cell subsets, defined as shown in (A,B), after stimulation of PBMCs with the indicated TLR ligands. Triplicates cultures from three different animals are shown. (D) Proliferation of FACS-sorted B cell subsets after stimulation for 3 days was determined using 3H-thymidine incorporation. P1 was defined and sorted as CD21+IgM+ subset, P2 as IgG+ subset, P3 as CD21IgM+CD11R1+ subset, and P4 as CD21IgM+CD11R1 subset (for gate definition see Figure 2). (C,D) Mean values are indicated by horizontal bars. Error bars show SDs. Statistical significance was calculated using ANOVA followed by Tukey’s test.