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. Author manuscript; available in PMC: 2016 Jul 25.
Published in final edited form as: Nat Med. 2015 Sep 21;21(10):1163–1171. doi: 10.1038/nm.3952

Figure 2.

Figure 2

MYC and inflammation are key tumorigenic drivers of PDAC that are inhibited by JQ1 treatment and BET protein inhibition. (a) Gene set enrichment analysis (GSEA) in JQ1-treated compared to vehicle-treated Kras;p53 mouse primary PDAC cells. (b) Quantification of spontaneous PanIN lesions formed in 6-month-old Kras (n = 8) and Kras;Myc (n = 8) mutant mice. The grade of lesions is indicated. **P < 0.01; ***P < 0.001 (two-tailed unpaired Student’s t-test). Data are represented as mean ± s.e.m. (c) GSEA from data sets comparing vehicle- to JQ1-treated Kras;p53 mouse primary PDAC cells (see also Supplementary Fig. 5g). (d) Relative serum concentration of inflammatory cytokines in the serum of Kras;p53 mutant mice treated with vehicle control or JQ1 (n = 3 for each experimental group) and wild-type (WT) animals, (e) Quantitative RT-PCR analysis of IL6 and IL1a mRNA expression in patient-derived PDAC xenografts following treatment with JQ1 or vehicle control (see Fig. 4e) (six biological replicates for each experimental condition). **P < 0.01; ***P < 0.001 (two-tailed unpaired Student’s t-test). Data are represented as mean ± s.e.m. (f) Effects of BRD4 inhibition via shRNA or JQ1 treatment on IL6 levels in the conditioned medium of human PDAC CFPac1 cells and for MYC and STAT3 as assessed by immunoblot. **P < 0.01; ***P < 0.001; n.s., not significant (two-tailed unpaired Student’s t-test). Data are represented as mean ± s.e.m. (g) Schematic representation of the human IL6 promoter (UCSC Gene Browser) with integrated epigenetic regulation marks and RNA polymerase II binding (POL2RA, green bar) (ENCODE). Localization of primers used for chromatin immunoprecipitation analysis is indicated by A–F. (h) Chromatin immunoprecipitation analysis of BRD4 at the IL6 promoter in CFPac1 cells treated with vehicle control (−) or JQ1 (+). The data are plotted relative to the values obtained with IgG control antibodies. **P < 0.01; ***P < 0.001 (two-tailed unpaired Student’s t-test). Data are represented as mean ± s.e.m. (i) Schematic of the caerulein pancreatitis– induced preneoplastic (PanIN) lesion formation protocol for the rescue experiment with exogenous IL6 injection upon JQ1 treatment. (j) Top, representative pancreata images (of n = 5 each, scale bars, 1 cm) and HE staining (scale bars, 100 µm) in Kras mutant mice in the four experimental conditions indicated (IHC and immunoblot biopsy analysis shown in Supplementary Fig. 7a,b). Bottom, quantification of MUC5AC-positive lesions in caerulein-treated pancreata from each experimental group (the control group was injected with vehicle control) (n = 5 each; IHC staining on Supplementary Fig. 7a). ***P < 0.001; n.s., not significant (two-tailed unpaired Student’s t-test). Data are represented as mean ± s.e.m.