Figure 1. Blimp-1 and c-Maf segregate with IL-10 expression in Th1 cells. (A) Heat map representation of relative expressions of transcription factors >2× up-regulated in in vitro generated IL-10–producing versus nonproducing Th1 cells. Cells were FACS-sorted according to their IL-10 secretion after IL-10 cytokine secretion assay 5 d after activation with anti-CD3/anti-CD28 MACSi Beads under Th1 skewing conditions. Probes were ranked by the difference (log2-fold) between results obtained from IL-10+Th1 cells and IL-10−Th1 cells (FC > 2). (B) Relative Prdm1 and Maf expression in FACS-sorted IL-10–secreting (IL-10+) and nonsecreting (IL-10−) Th1 cells as determined by qPCR. Cells were generated and sorted similarly to A. From left to right: ***, P = 0.0006; *, P = 0.0189. (C) Blimp-1 and c-Maf protein level in IL-10–producing (IL-10:GFP+) and nonproducing (IL-10:GFP−) in vitro generated Th1 cells from IL10:GFP reporter mice 72 h after activation as determined by immunoblot. Indication of protein size is in kD. (D) Relative Il10, Prdm1, and Maf expression time course in indicated in vitro generated Th cell subsets measured by qPCR. (E) Blimp-1 and c-Maf protein expression in indicated in vitro generated Th cell subsets as determined by immunoblot 72 h after T cell priming. Indication of protein size is in kD. (F) Comparison of Prdm1 and Maf expression, measured by qPCR, in IL-10/GFP+ versus IL-10/GFP− in vitro generated Th1, Th2, and Th17 cells. Cells were FACS-sorted 72 h after starting the Th cell cultures. From left to right: **, P = 0.0089; *, P = 0.037; *, P = 0.048; *, P = 0.043. Data are representative of at least 2 independent experiments (error bars, mean ± SEM).
Figure 1.

Blimp-1 and c-Maf segregate with IL-10 expression in Th1 cells. (A) Heat map representation of relative expressions of transcription factors >2× up-regulated in in vitro generated IL-10–producing versus nonproducing Th1 cells. Cells were FACS-sorted according to their IL-10 secretion after IL-10 cytokine secretion assay 5 d after activation with anti-CD3/anti-CD28 MACSi Beads under Th1 skewing conditions. Probes were ranked by the difference (log2-fold) between results obtained from IL-10+Th1 cells and IL-10−Th1 cells (FC > 2). (B) Relative Prdm1 and Maf expression in FACS-sorted IL-10–secreting (IL-10+) and nonsecreting (IL-10−) Th1 cells as determined by qPCR. Cells were generated and sorted similarly to A. From left to right: ***, P = 0.0006; *, P = 0.0189. (C) Blimp-1 and c-Maf protein level in IL-10–producing (IL-10:GFP+) and nonproducing (IL-10:GFP−) in vitro generated Th1 cells from IL10:GFP reporter mice 72 h after activation as determined by immunoblot. Indication of protein size is in kD. (D) Relative Il10, Prdm1, and Maf expression time course in indicated in vitro generated Th cell subsets measured by qPCR. (E) Blimp-1 and c-Maf protein expression in indicated in vitro generated Th cell subsets as determined by immunoblot 72 h after T cell priming. Indication of protein size is in kD. (F) Comparison of Prdm1 and Maf expression, measured by qPCR, in IL-10/GFP+ versus IL-10/GFP− in vitro generated Th1, Th2, and Th17 cells. Cells were FACS-sorted 72 h after starting the Th cell cultures. From left to right: **, P = 0.0089; *, P = 0.037; *, P = 0.048; *, P = 0.043. Data are representative of at least 2 independent experiments (error bars, mean ± SEM).

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