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. 2022 Feb 20;12(2):e654. doi: 10.1002/ctm2.654

FIGURE 4.

FIGURE 4

PTGS2 antisense NF‐κB1 complex‐mediated expression regulator RNA (PACERR) binds to the promoter region of prostaglandin‐endoperoxide synthase 2 (PTGS2) and positively regulates PTGS2 expression. (A) Linear regression and Spearman correlation analysis of the relative expression of PACERR and PTGS2 in tumour‐associated macrophages (TAMs) isolated from pancreatic ductal adenocarcinoma (PDAC) tissues (n = 9). (B) Fluorescence in situ hybridisation (FISH) of PACERR (green) in THP‐1‐derived TAMs. DAPI staining (blue) shows the nuclei. Scar bar: 10 μm. (C) Graphical scheme describing the workflow of chromatin isolation through RNA purification with sequencing (ChIRP‐seq) experiment. (D) Genome browser snapshots of ChIRP‐seq signals for the genomic regions near PTGS2 and PACERR in THP‐1‐derived TAMs (shNC/shPACERR, stimulated with phorbol 12‐myristate 13‐acetate [PMA] and co‐cultured with PANC‐1). shNC means that cells were transfected in negative control plasmids. The PACERR binding sites are shown as dashed lines. The histogram displays ChIRP‐seq signals (fold enrichment over input) of PACERR near PTGS2 promoter regions in shNC and shPACERR THP‐1‐derived TAMs. (E) PTGS2 protein and mRNA expression examined by Western blot assay and qPCR analysis, respectively, in THP‐1‐derived TAMs (shNC/ shPACERR, stimulated with PMA and co‐cultured with PANC‐1)