Fig. 8

Tumoral c-Jun binds to the promoter of stemness and iron metabolism related genes to up-regulate their transcription. A Venn diagram of stemness genes and iron metabolism genes from RNA-sequencing data of PDX1#-vector/c-Jun-OE. B RNA-seq expression level for a panel of stemness and iron metabolism gene in the PDX1#-vector/c-Jun-OE cell lines. C-F Q-PCR for c-Jun/SOX9/SOX2/NANOG/OCT4/FTH1/FTL/ TFRC were performed in indicated cell lines. G Western blot for c-Jun/SOX9/SOX2/NANOG/OCT4/FTH1/FTL/ TFRC were performed in indicated cell lines. H ChIP-seq on PDX1#-c-Jun cell lines were performed. Representative ChIP-seq input, H3K27ac, and c-Jun peaks for stemness genes (SOX9/SOX2/OCT4) and iron metabolism genes (FTH1/FTL/TFRC). I-J Ch-IP assay was performed to validate binding of c-Jun on promoter of indicated genes in PDX1# cell lines. K-L Dual luciferase assay was performed to determine the promoter activity in PDX1#-vector/c-Jun-OE cell lines. Renilla luciferase activity was used as internal control. All experiments were repeated three times independently. Paired Student’s t-test were used for in vitro experiments