Fig. 4

Three-dimensional epigenome reprogramming upregulates GATM expression promoting PDAC metastasis. A In situ Hi-C maps and histone ChIP-seq tracks surrounding the Gatm gene locus. The enhancer-promoter loop was showed by black circle. B ChIP-qPCR showing H3K27ac enriched to Gatm promoter and candidate enhancers. Heatmap visualization of ChIP-qPCR data that was divided into GATM low expression group and high expression group. C 3C-PCR showing the interaction between the candidate enhancers and Gatm promoter. D-E RT-qPCR showing Gatm expression in PANC-1-IN-dCas9-KRAB cells and Capan-1-dCas9-KRAB cells which were transfected by phU6 control and vectors carrying specific sgRNAs. (F-G) Western blot and transwell migration assay showing GATM expression and migration ability of PANC-1-IN-dCas9-KRAB cells. 200 × magnifying power. The groups named g10947, g10948 and g10949 stably expressed sgRNAs targeting Enh#4, 5 and 6 respectively. H-I Representative IVIS bioluminescence images and quantification of PDAC liver metastasis in splenic injection NPG mouse model. (h) animal level images; (i) dissected liver images. Data in (B-C), (D-E) and (G-I) are presented as mean ± SEM by 2way ANOVA (Tukey’s multiple comparisons test), unpaired t tests and ordinary one-way ANOVA (Tukey’s multiple comparisons test) respectively. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001