Fig 5. PGE2 inhibits c-Maf in CD4 T cells differentiated in the presence of IL-27.

(A) Naïve CD4⁺CD62L⁺ cells were stimulated in the presence of IL-27 and PGE2. RNA was collected at 24, 48 and 72 hr and analyzed by RT-PCR for Maf and Ahr expression. (B) Naïve CD4⁺CD62L⁺ cells were stimulated in the presence of IL-27 and PGE2 or dbcAMP. Cells were collected on day 3 and analyzed by RT-PCR for Maf expression. (C) Splenocytes from il10^gfp mice were stimulated with 3 μg/ml anti-CD3 and 1 μg LPS in the presence of IL-27 and PGE2. RNA was collected on day 3 and Maf expression was analyzed by RT-PCR. (D) Naïve CD4⁺CD62L⁺ cells were stimulated in the presence of IL-27 and PGE2 or dbcAMP. Cells were collected on day 3 and analyzed by FACS for intracellular c-Maf within the whole cell population (upper panel) and within CD49b⁺LAG-3⁺ populations (lower panel). Graphical representation of data is on the right. (E) Naïve CD4⁺CD62L⁺ cells were stimulated as in D. Cells were collected on day 3 and analyzed by FACS for intracellular Egr-2 within the whole cell population (upper panel) and within CD49b⁺LAG-3⁺ populations (lower panel). (F) Naïve CD4⁺CD62L⁺ cells were stimulated as in D. Cells were collected on day 3 and analyzed by FACS for intracellular Blimp-1 within the whole cell population and within CD49b⁺LAG-3⁺ populations. Data are representative of two to three independent experiments. Each sample was tested in duplicate and results represent means ± SD. Significance was evaluated by one-way ANOVA and * symbolizes significance of experimental condition versus IL-27 control (B, D, E); *P<0.05, **P<0.01, ***P<0.001.