Fig. 4

YTHDF2 mediates NEDD4 mRNA expression A RNA pull down with NEDD4 RNA detected by YTHDF2 antibody. B The NEDD4 m6A alteration caused by YTHDF2 in CFPAC-GM and Colo357-GM cells was shown by RIP-qPCR analysis. C In YTHDF2-silenced CFPAC-GM and Colo357-GM cells, the relative activity of the WT or Mut luciferase reporters of NEDD4 was measured and normalized to the negative control groups. D RNA stability assay showed that YTHDF2 can maintain NEDD4 mRNA stability. E YTHDF2 regulates NEDD4 expression in CFPAC-GM and Colo357-GM cells. F The chemosensitivity of CFPAC-GM and Colo357-GM cells to gemcitabine was assessed using CCK-8 toxicity tests. The IC50 of cells transfected with shYTHDF2 and NEDD4 vector were analyzed. G, H Silencing YTHDF2 or overexpressing NEDD4 in CFPAC-GM cells influenced pancreatic tumor growth in nude mouse models treated with gemcitabine. Group A: cells transfected with shYTHDF2-NC. Group B: cells transfected with shYTHDF2-1. Group C: cells transfected with both shYTHDF2-1 and NEDD4 overexpression plasmid. Group D: cells transfected with shYTHDF2-1 and NEDD4 control vector