Supplementary Figure 4: Regulation of Treg cells in mixed chimeras and Ptenfl/flFoxp3-Cre Ifng−/− mice.
From: Treg cells require the phosphatase PTEN to restrain TH1 and TFH cell responses
(a) Sublethally irradiated Rag1−/− mice were reconstituted with a 1:1 mix of CD45.1+ BM and either CD45.2+ WT or Ptenfl/flFoxp3-Cre BM cells. Following reconstitution, the mixed chimeras were analyzed for intracellular staining of IFN-γ in CD8+ T cells. (b) Cytokine production of IFN-γ, IL-17 and IL-4 by CD4+ T cells from WT, Ptenfl/flFoxp3-Cre, Ifng−/− and Ptenfl/flFoxp3-Cre Ifng−/− mice after in vitro stimulation for 4 h. (c) PNA staining of spleen sections (magnification, x2; scale bars, 2 mm). (d) Immunofluorescence of MLN sections for the staining of CD3 (red), PNA (green) and IgD (white) (scale 500 μm). Right, quantification of germinal center area. Data are representative of three independent experiments (a,b) and one experiment (c,d; n = 3 WT, 6 Ptenfl/flFoxp3-Cre, 2 Ifng−/− and 3 Ptenfl/flFoxp3-Cre Ifng−/− mice). Data are mean ± s.e.m. Numbers indicate percentage of cells in quadrants or gates.
