EBV-miR-BART1-5P directly targets cellular AMPKα1 gene.

(A) EBV-miR-BART1-5P and its putative binding sequences in the 3’UTR of AMPKα1. Mutation was generated in the complementary site that binds to the seed region of BV-miR-BART1-5P. (B) HEK293T cells were cotransfected with EBV-miR-BART1-5P mimic or NC and luciferase reporter carrying either the predicted miRNA target site in AMPKα1 3’UTR (wt) or its corresponding mutant (mut). Inhibitor of EBV-miR-BART1-5P by anti-miR and anti-control (anti-c) was also performed. The data were shown as the mean ± s.e.m. (***PC) Western blotting analysis of endogenous AMPKα1 protein expression levels in CNE1 and HONE1 cells treated with EBV-miR-BART1-5P mimic or inhibitor. β-actin was used as a loading control. (D) AMPKα1 expression was detected using immunohistochemistry assay in tumour tissues derived from tumorigenesis in nude mice models. Magnification, ×400. (E) The levels of AMPKα1 was evaluated by qPCR in NPC and NP tissue specimens (with clinical stage and T stage). Normalized to GAPDH. (F) The correlation between EBV-miR-BART1-5P and AMPKα1 expression levels was calculated. The data were shown as the mean ± s.e.m. (*P<0.05, **P<0.01 and ***P<0.001).