Fig. 7: MIR181A1HG is regulated by the NFATC2-SOCS3 complex in GC cells. | Oncogene

Fig. 7: MIR181A1HG is regulated by the NFATC2-SOCS3 complex in GC cells.

From: Long non-coding RNA-MIR181A1HG acts as an oncogene and contributes to invasion and metastasis in gastric cancer

Fig. 7

A After transfection with siRNAp of TFs or scrambled-siRNA (Scr-siRNA) in AGS and MKN-74 cells, MIR181A1HG expression was detected using qRT-PCR. *, P > 0.05, and ***, P < 0.01. siRNAp, siRNA pool; TFs, Transcription factors. B The GC cells transfected with NFATC2 or Vector, expression levels of MIR181A1HG, miR-181a-5p or miR-181b-5p were detected by qRT-PCR. **, P < 0.05, ***, P < 0.01, and ****, P < 0.001. C The interactions beween NFATC2 and SOCS3 in AGS and MKN-74 cells were assessed by Co-IP assays using antibodies against Flag-tag (SOCS3). D1 Illustration of CRISPR interference. D2 MIR181A1HG, miR-181a-5p or miR-181b-5p expression were assayed by qRT-PCR with CRISPR interference. (E) The transcriptional factor NFATC2 binding motif was predicted using Jaspar database. F Schematic representation of the promoter region of MIR181A1HG (MIR181A1HGp). G ChIP assay demonstrated the direct binding of NFATC2 to the site 1 of MIR181A1HGp in AGS and MKN-74 cells. H WT or MT MIR181A1HG promoter constructs were co-transfected with NFATC2 or Vector, and the relative luciferase activity was determined. WT: wildtype; MT: mutated. *, P > 0.05, and ****, P < 0.001. I, J MIR181A1HG expression were detected after transfection by qRT-PCR. **, P < 0.05, and ***, P < 0.01. K, L The MIR181A1HG promoter construct was co-transfected with SOCS3 or/and NFATC2 plasmids or siRNAs or corresponding control. Luciferase assays were performed. ***, P < 0.01, and ****, P < 0.001. M, N Invasion (M) and migration (N) assays were conducted and quantitative results are shown in GC cells. **, P < 0.05, ***, P < 0.01, and ****, P < 0.001. (O) Wound closure percentages in GC cells are shown. ***, P < 0.01, and ****, P < 0.001.

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