Fig. 3.
Role of hcIE in the Bcl6 suppressive function for TH2 cytokine gene loci. (A) A conserved sequence [positions +1,180 to +1,401 relative to the transcription start site; Mouse Genome Informatics (MGI) accession no. 96556] in the HS2 region of mouse hcIE is shown along with human hcIE. Conserved sequences between mice and humans are indicated by shaded boxes. (B) Cytokine production (EP and LP) by WT and hcIE-KO TH2 cells estimated after restimulation with anti-CD3 Abs for 48 h. (C and D) Analysis of binding levels of (C, Upper) Bcl6, (C, Lower) STAT5, and (D) GATA3 by ChIP–quantitative PCR for hcIE-KO and WT TH2 cells. (E and F) Effect of Bcl6 on cytokine gene expression estimated in EPs and LPs after restimulation for 24 h and acetylation of histone H3 at K9 and K14 in EPs at rest with/without Bcl6 inhibitor in hcIE-KO and WT TH2 cells. Data are shown as means ± SDs. Data are representative of (B) four, (C) eight, (D and E) six, or (F) seven independent experiments with (B) n = 4–5, (C) n = 7–8, (D) n = 5–6, (E) n = 6–8, or (F) n = 6–7 samples in each experiment. ND, not detected; NS, not significant. *P < 0.05; **P < 0.01.