To further dissect whether loss of p16 upregulates NOX4 expression via the Rb-E2F pathway in pancreatic cancer cells, we performed an immunoblotting analysis, which showed that Rb was strongly phosphorylated at sites S795 and S807/811 and that the expression level of nuclear E2F1 was increased in HPNE/KrasG12V/shp16 cells (Fig. 5a). This evidence concerns the gene CDKN2A and familial pancreatic carcinoma.