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. 2025 Jan 6;16:335. doi: 10.1038/s41467-024-55330-7

Fig. 3. JUNB-HDAC1 complex represses inflammatory signals and cJUN.

Fig. 3

a Heatmap showing expression of cytokines present in the core enrichment of the gene sets shown in Supplementary Fig. 3a–d, for JUNB silencing (siJUNB; n = 3 biological replicates) versus control siRNA (siCtrl; n = 2 biological replicates) in CAPAN1 cells. Cell color indicates z score. b qRT-PCR analysis for indicated target genes in siJUNB conditions (red), normalized to siCtrl (gray), in CAPAN1. Relative mRNA expression with mean ± s.d. shown. n = 3 biological replicates. Two-tailed Student’s t-test with Welch’s correction. Coverage of JUNB ChIP-seq data in CAPAN121, as well as publicly available H3K27ac25 data, for loci of cJUN (c), IL1A/B (d), and CXCL9/10/11 (e). ChIP-qPCR validation regions are indicated. f Gene set enrichment analysis for curated signatures (C2) of the Molecular Signature Database (MSigDB) for siJUNB versus siCtrl in CAPAN1 cells. Normalized enrichment score (NES) and FDR q value are indicated. Immunoblot for JUNB, HDAC1, and β-actin after JUNB pulldown, IgG isotype control or input in CAPAN1 (g) and CAPAN2 (h). n = 3 biological replicates. i, j, ChIP-qPCR for regions indicated in (c–e), showing signal relative to input for JUNB (i) and HDAC1 (j) pulldown with mean ± s.d. and average IgG isotype control. n = 3 biological replicates. k, l, ChIP-seq analysis for JUNB in control cells (as in c–e) and HDAC1 with siJUNB or siCtrl. k Overlap of JUNB binding regions and regions where HDAC1 is significantly lost upon siJUNB (“HDAC1_DOWN”). l, GREAT analysis of the overlapping regions of (k) with –log10(Padj) for binomial test indicated. Hallmark (H) and C2 signatures of MSigDB are shown. Representative immunoblot for JUNB, cJUN, CCL2, and β-actin in HPAF-II (m) and CFPAC-1 (n) after siJUNB or siCtrl. n = 3 biological replicates. o Dual-luciferase reporter assay for cJUN promoter firefly luciferase (Luc) constructs in CAPAN2 cells with varying concentrations of JUNB overexpression plasmids (or EV controls). Relative Luc activity to Renilla luciferase control with mean ± s.d. shown. One-way ANOVA. n = 3 biological replicates. Immunoblot for JUNB, cJUN, and β-actin in CAPAN1 (p) and CAPAN2 (q) cells with overexpression of cJUN (cJUN-OE) or empty vector (EV) control. n = 3 biological replicates. Source data are provided as a Source Data file.