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Fig. 1 | Journal of Translational Medicine

Fig. 1

From: Amelioration of autoimmune arthritis by adoptive transfer of Foxp3-expressing regulatory B cells is associated with the Treg/Th17 cell balance

Fig. 1

Foxp3 is expressed by LPS and anti-IgM in mouse B cells. Purified CD19+ B cells were isolated from spleens of 6- to 8-week-old DBA/1J mice. CD19+ primary B cells isolated from splenocytes of wild type mice or A20 (mouse B cell line) were stimulated with 10 µg/ml LPS, or 10 µg/ml anti-IgM for 72 h. Foxp3 a mRNA and b protein levels were determined by RT-PCR and western blot, respectively. The optical densities of Foxp3 bands were normalized to those of β-actin. The data represent one of three similar experiments. Bar graphs show mean ± SD. Data are representative of three independent experiments. Significant differences between means of cells cultured in media alone and stimulated cultures are indicated: *p < 0.05, **p < 0.01 compared with untreated cells. c Whole splenocytes were stained with CD19-FITC mAb before permeabilization and stained using a Foxp3-PE mAb. Numbers indicate Foxp3+ cell frequencies within CD19+ B cell subsets. The graph on the right represents the absolute cell numbers of Foxp3+ Breg cells. Data are representative of three independent experiments. d Confocal microscopic analysis of cultured CD4-deplected splenocytes for Foxp3 (red, PE), and CD19 (green, FITC). Representative images after LPS treatment are shown. Original magnification, ×400

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