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. 2018 Mar 9;11:367–381. doi: 10.1016/j.omtn.2018.03.002

Figure 3.

Figure 3

P62 Regulates IGFII Transcriptional Activity in the Mesenchymal Stem Cells with TNF-α Treatment

(A) RNA immunoprecipitation (RIP) with anti-CTCF followed by RT-PCR with CUDR mRNA primers in the mesenchymal stem cells treated with TNF-α and transfected with pCMV6-AC-GFP (GFP-control), pCMV6-AC-GFP-P62 (GFP-P62), pGFP-V-RS (RNAi control), and pGFP-V-RS–P62 (P62 RNAi). IgG RIP was used as negative control. CUDR RNA as INPUT is shown. (B) Chromosome conformation capture (3C)-chromatin immunoprecipitation (ChIP) with anti-CTCF and anti-RNA polII in the mesenchymal stem cells treated with TNF-α and transfected with pCMV6-AC-GFP, pCMV6-AC-GFP-P62, pGFP-V-RS, and pGFP-V-RS-P62. The PCR analysis is applied for detecting IGFII H19ICR-IGFII DMR2 coupling product using H19ICR and IGFII DMR2 primers. The H19ICR and IGFII DMR2 as INPUT are shown. (C) 3C-ChIP with anti-CTCF and anti-RNA polII in the CUDR knockdown mesenchymal stem cells transfected with pCMV6-AC-GFP, pCMV6-AC-GFP-P62, pGFP-V-RS, and pGFP-V-RS-P62. The PCR analysis is applied for detecting IGFII H19ICR-IGFII DMR2 coupling product using H19ICR and IGFII DMR2 primers. The H19ICR and IGFII DMR2 as INPUT are shown. (D) IGFII promoter luciferase activity assay in in the mesenchymal stem cells treated with TNF-α and transfected with pCMV6-AC-GFP, pCMV6-AC-GFP-P62, pGFP-V-RS, and pGFP-V-RS-P62. Each value was presented as mean ± SEM. **p < 0.01.