(A) Upper panels: The inhibitor's effect on NF-κB-regulated genes—defined as per http://www.bu.edu/nf-kb/gene-resources/target-genes. Left, relevant transcripts from pancreatic CD45+ cells of NOD mice treated long- or short-term with I-BET151 or DMSO. Red: over-represented; blue: under-represented. 2 wk: long-term, treatment as per Figure 1B; 48 hr: short-term, treatment with a single 10 mg/kg dose and analyzed 48 hr later. Right, signaling pathways represented by the enriched or impoverished transcripts in the data to the left, via Ingenuity pathway analysis (www.ingenuity.com). Lower panels: Gene sets corresponding to the TNFα-induced canonical NF-κB pathway (Schaefer et al., 2009) or the PPAR and LXR pathways (http://www.genome.jp/kegg/pathway/hsa/hsa03320.html) were retrieved from the Broad Institute's Molecular Signatures Database (http://www.broadinstitute.org/gsea/msigdb), and their expression levels in CD45+ cells from pancreas of I-BET151- or vehicle-treated mice plotted. (B) 12-week-old NOD mice were injected once ip with BAY 11–7082 (10 mg/kg), sacrificed 24 hr later, and CD45+ cells from the pancreas isolated and transcriptionally profiled. A volcano plot comparing treatment with BAY 11–7082 and DMSO, with genes >twofold increased (in red) or decreased (in blue) by I-BET151 treatment (pooled from all time-points of Figure 3A,B) superimposed. (C) Effect of a single dose of 10 mg/kg BAY 11–7082 on insulitis in 12-week-old NOD mice, analyzed 24 hr after injection. Left: insulitis scores. Right: summary data for the fraction of islets with no infiltrate. Grey, peri-insulitis; Black, insulitis. (D) Suppression of in vitro T cell proliferation by cell populations isolated from the pancreas of I-BET151- or DMSO- treated mice (as per Figure 1B). The CD11b+CD11c− (top), CD11b−CD11c+ (middle) and TCRβ+CD4+CD25+ (bottom) fractions of CD45+ cells were sorted. To the left are representative plots of CFSE dilution; to the right are summary data quantifying division indices (see ‘Materials and methods’ for details). p values in A and B are from the chi-squared test, and in C and D are from the Student's t test.