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Fig. 8 | Journal of Hematology & Oncology

Fig. 8

From: LncRNA-PACERR induces pro-tumour macrophages via interacting with miR-671-3p and m6A-reader IGF2BP2 in pancreatic ductal adenocarcinoma

Fig. 8

LncRNA-PACERR cooperates with IGF2BP2 to regulate KLF12 and c-myc in an m6A-dependent manner. A, B qRT-PCR analysis of the LncRNA-PACERR, KLF12 and c-myc transcript levels in the LncRNA-PACERR KD (A) and IGF2BP2 KD (B) THP-1 derived TAMs (n = 3). C, D Half-life of KLF12 after treatment with 5 μmol/L actinomycin D for the indicated times in the IGF2BP2 KD THP-1 derived TAMs with ectopically expressed LncRNA-PACERR (C) and in the LncRNA-PACERR KD THP-1 derived TAMs with ectopically expressed IGF2BP2 (D). E, F Falf-life of c-myc after treatment with 5 μmol/L actinomycin D for the indicated times in the IGF2BP2 KD THP-1 derived TAMs with ectopically expressed LncRNA-PACERR (E) and in the LncRNA-PACERR KD THP-1 derived TAMs with ectopically expressed IGF2BP2 (F). G RIP qRT-PCR showing the enrichment of m6A modification in the KLF12 3′ UTR/5′ UTR and c-myc CRD regions in the METTL14 KD THP-1 derived TAMs (n = 3). H, I RIP qRT-PCR detecting the enrichment of IGF2BP2 (H) and biotin-labelled LncRNA-PACERR (I) in the KLF12 3′ UTR and c-myc CRD in LncRNA-PACERR KD (H) and IGF2BP2 KD (I) THP-1 derived TAMs (n = 3). J Schematic representation of wild-type (WT) and mutated (MUT; GGAC to AAGT) KLF12 3′ UTR of the pmirGLO vector. K, L RIP qRT-PCR detection of the enrichment of IGF2BP2 (K) and m6A (L) in the KLF12 3′ UTR WT and MUT luciferase reporters in the LncRNA-PACERR and IGF2BP2 OE cells (n = 3). M Relative luciferase activity levels of KLF12 3′ UTR WT and MUT reporters in the LncRNA-PACERR and IGF2BP2 OE cells (n = 3). N Proposed model demonstrating a positive feedback loop between LncRNA-PACERR and KLF12 in TAMs to promote proliferation and migration in PDAC. The expression of LncRNA-PACERR is activated by EP300-mediated H3K27 acylation. LncRNA-PACERR exerts its pro-tuomour function by regulating miR-671-3p/KLF12/AKT/c-myc axis and sequestering IGF2BP2

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