Fig. 5: IL-6 transcriptionally elevated DLGAP1-AS1 expression in HCC cells through JAK2/STAT3 signaling pathway. | Cell Death & Disease

Fig. 5: IL-6 transcriptionally elevated DLGAP1-AS1 expression in HCC cells through JAK2/STAT3 signaling pathway.

From: Long non-coding RNA DLGAP1-AS1 facilitates tumorigenesis and epithelial–mesenchymal transition in hepatocellular carcinoma via the feedback loop of miR-26a/b-5p/IL-6/JAK2/STAT3 and Wnt/β-catenin pathway

Fig. 5

a The STAT3 motif predicted from JASPAR database. b Luciferase reporter assay demonstrated luciferase activities of various truncated reporters in Hep G2 cells, determining the region of DLGAP1-AS1 promoter on which STAT3 could bind to mediate transcriptional activation. c ChIP assay was performed using the STAT3 antibody to demonstrate the enrichment of the STAT3-binding region of DLGAP1-AS1 promoter in Hep G2 and SNU-387 cells. d WB analysis evaluated phosphorylated STAT3 levels in HCC cell lines and in normal cells. (e) qRT-PCR evaluated the efficiency of STAT3 overexpression in SNU-387 cells. f WB analysis evaluated the efficiency of STAT3 overexpression and the activating effect of IL-6 on JAK2/STAT3 pathway in SNU-387 cells, while JAK2/STAT3 pathway inhibitor Cucurbitacin I was applied so that the IL-6-induced activation of JAK2 and STAT3 was reversed. g Luciferase reporter assay was performed in SNU-387 cells to verify that STAT3 interacted with DLGAP1-AS1 promoter at the predicted binding motif. The interaction was enhanced by IL-6 treatment and repressed by Cucurbitacin I treatment. h DLGAP1-AS1 expression levels influenced by STAT3, IL-6, and Cucurbitacin I were assessed using qRT-PCR. All data are presented as the mean ± SD of three independent experiments. **p < 0.01.

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