CD44v6+ Hepatocellular Carcinoma Cells Maintain Stemness Properties through Met/cJun/Nanog Signaling

Figure 4

The transcription factor, cJun, mediates HGF/MET signaling and regulates Nanog expression. (a) STRING (https://string-db.org/cgi/input.pl) analysis showing functional association networks of cJun and Met-correlated genes. Gene Set Enrichment Analysis (GSEA) was used to analyze the expression of Met associated with the transcription factor cJun. (b) Immunoblotting analysis results showed that the expression of Nanog recovered after knockout of Met and transferred to cJun in the meantime. β-Actin was used as a normalized control. (c) Immunoblotting analysis showed the expression of cJun was barely changed when we overexpressed the Nanog. β-Actin was used as a normalized control. (d) Luciferase reporter assay analysis of Nanog promoter luciferase reporters in SNU398 cells before luciferase detection. It showed that there is a cJun binding site in the 0–2000 bp upstream regions of Nanog promoter. (e) The prediction of the binding sites of cJun to DNA in Jaspar Database (http://jaspar.genereg.net). And the schematic diagram shows three luciferase reporters cover different DNA sequences of Nanog promoter region. (f) Luciferase reporter assay analysis of three Nanog promoter luciferase reporters in SNU398 cells transfected with cJun or pcDNA. , -test. (g) ChIP assay showed that primer2 is the exact binding site of cJun. .

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