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. 2018 Jun 21;13(6):e0199332. doi: 10.1371/journal.pone.0199332

Fig 4. Wnt3a stimulation increment proliferation of B-1 cells in response of IL7.

Fig 4

Purified B-1 cells from C57BL/6 mice were treated with Wnt3a (100ng/ml), IL-7 (50ng/ml) and Wnt3a+IL7 during 3 days. Non-treated cells (NT) were used as control group. A) Contour plots analysis of expression of CD19 and IL7R by non-treated B-1 cells or Wnt3a-treated B-1 cells. Histograms show the proliferation (CFSE decay) of IL7R+(blue) or IL7R-(red) B-1 cells in control group or Wnt3a group. The maximum of CFSE staining cells in time zero were considered to determine the region gate of non-proliferative cells. To measured the decay of fluorescence, which is not related to proliferation, B-1 cells cultured in a RPMI medium only was used. The decay of fluorescence in these samples was subtracted to the decay of fluorescence in the experimental groups to determine the gate region of fluorescence cells and also MFI. B) Absolute number of B-1 cells IL7R- or IL7R+ cells from non-treated group (NT) or Wnt3a treated group (Wnt3a). C) Proliferation of IL7R- or IL7R+ B-1 cells from non-treated group (NT) or Wnt3a treated group (Wnt3a) measured by decay in the MFI value. D) Histograms of CFSE decay (proliferation) of B-1 cells in the different groups: NT (non-treated), Wnt3a, IL-7, Wnt3a+IL7. E) Proliferation of B-1 cells was represented by MFI value in different conditions: NT (non-treated), Wnt3a, IL-7, Wnt3a+IL7. ***p<0,001, **p<0,01 and *p<0,05 (One way ANOVA). Data from a representative of 3 independent experiments performed in A,B and C and 2 independent experiments performed in D and E. Each experiment was performed using 3 biological samples.