Fig. 7

Tumoral c-Jun functionally increases stemness and iron metabolism pathways in PDAC. A Top enriched pathways in PDO4#-vector/c-Jun-OE and primary cell line PDX1#-vector/c-Jun-OE based on Gene Set Enrichment Analysis on RNA-sequencing data. NES, normalized enrichment score, Norm. p, normalized P value. B Enrichment score profile of the stemness (left) and iron homeostasis pathway (right) in GSEA. C-D Sphere formation assays were performed in indicated cell lines. E–F In vitro limited dilution assays were performed in indicated cell lines. G-H In vivo limited dilution assays for PDX1# cell lines were performed. Representative tumor images (G) and tumor incidence/CSC probabilities (H) were shown. I Schematic illustration of the experimental procedure for the in vitro transferrin uptake in cancer cells. J-K One representative lane from the 3D reconstructions of representative fields showing transferrin (red) in vector or scramble cells (blue) and c-Jun-OE or c-Jun-KD cells (green) and quantification of fluorescent-transferrin as measured on the surface of the 3D reconstruction. L-M Iron ion content was determined in indicated cell lines (N–O) Relative libel iron pool (LIP) in indicated cell lines were determined by FCM. MFI of calcein in cells with or without defetoxamine (DFO) treatment were analyzed and ΔMFI were calculated as libel iron pool (LIP). All experiments were repeated three times independently. Paired Student’s t-test were used for in vitro experiments