Fig. 4: Accessibility of PD-L1 (CD274) and CD47 depended on JUN.
From: Activation of JUN in fibroblasts promotes pro-fibrotic programme and modulates protective immunity
a Heatmap demonstrating dynamic chromatin changes in fibrotic lung fibroblasts with (JUN-KO) or without (Control) JUN deletion and normal lung fibroblasts with (TetO-JUN Dox+) or without (TetO-JUN Dox−) JUN activation. b Representative genome browser tracks comparing ATAC-seq signal in fibrotic lung fibroblasts (with (JUN-KO) or without (Control) JUN-knockout) and also ChIP-seq signal in normal lung fibroblasts (with (TetO-JUN Dox+) or without (TetO-JUN Dox−) JUN overexpression) with A549, MCF-7, h1-hESC, HepG2 and K562 from published data at JUN, CD47 and CD274 loci. The red boxes highlight ATAC-seq and ChIP-seq peaks in the promoter sites of JUN, CD47 and CD274 (and enhancer is shown in green). We also compared our peaks with H3K4me3 or H3K27Ac (=histone mark for open chromatin), H3K9me3 or H3K27me3 (=histone mark for closed chromatin), ChIP-seq data generated from normal human-lung fibroblast is from published data, which highlighted the same areas respectively. c Gene expression changes in primary lung fibroblasts by comparing JUN knockout (KO) or overexpression (OE). Taqman assay were normalized to the value in JUN knockout. Data are expressed as mean ± SD from four experimental repeats. Two-tailed ratio paired t-test, **P < 0.01; ****P < 0.0001. d Representative flow cytometry histograms showing reduced expression of pJUN, PD-L1 and CD47 after JUN overexpression (OE) or knockout (KO). Yellow plot: JUN overexpression; Black plot: JUN knockout. e Vector maps of the control and CD47 enhancer constructs used to engineer reporter cell lines. f, g CD47 enhancer reporter assays demonstrating that doxycycline induced JUN expression initiated CD47 enhancer expression, which disappeared when JUN expression was turned off (f) or JUN was knocked out (g). Data are expressed as mean ± SD from three independent experiments, Ordinary one-way ANOVA (Tukey’s multiple comparisons test), n.s. non-significant; **P < 0.01; ***P < 0.001; ****P < 0.0001. See Supplementary Data 2 for statistical details. Source data are provided as a Source Data file.
