Fig. 6

KLF12 binds directly to LncRNA-PACERR and recruits EP300 to the promoter region of LncRNA-PACERR in a LncRNA-PACERR-dependent manner. A Association of KLF12 with the promoter region of LncRNA-PACERR in THP-1-derived TAMs (Crtl/KLF12 OE / shKLF12) analysed by ChIP-qPCR. B ChIP assays showed endogenous KLF12 binding to the LncRNA-PACERR gene promoter. C, D HEK293T cells were co-transfected with LncRNA-PACERR promoter–luciferase truncations and KLF12 plasmids, and the luciferase activity was determined using a dual luciferase reporter assay after 48 h. E Dual luciferase assay of HEK293T cells co-transfected with firefly luciferase constructs containing the wild-type or mutant KLF12 potential binding sites of LncRNA-PACERR promoter and KLF12 plasmids were performed. F Association of H3K27ac with the promoter region of LncRNA-PACERR in THP-1-derived TAMs (Crtl/ KLF12 OE/ shKLF12) analysed by ChIP-qPCR. G Results of coimmunoprecipitation (Co-IP) in THP-1-derived TAMs. Normal rabbit IgG was used as a negative control. H WB validation of KLF12 proteins pulled down with biotin-labelled LncRNA-PACERR is shown. I RNA immunoprecipitation (RIP) was performed using a KLF12-specific antibody. Eluted KLF12-binding RNAs were reverse transcribed, and qPCR was performed with primers specific for LncRNA-PACERR. Normal rabbit IgG (IgG) was used as a negative control. Data are shown as the results from three independent experiments. J Association of EP300 with the promoter region of LncRNA-PACERR analysed by ChIP-qPCR in THP-1-derived TAMs (shNC/ shLncRNA-PACERR + KLF12 OE/shKLF12 + LncRNA-PACERR OE/ shKLF12 + shLncRNA-PACERR)