Fig. 6

miR-18a reactivates EBV by targeting ATM. a DNA-PK inhibitor AZD 8055 increased EBV gene expression in Raji cells. Real-time PCR was used to measure the mRNA expression of EBV-related genes. GAPDH served as an internal control. b ATM is a potential target of miR-18a. I: Schematic representation of the 3’UTR of ATM. The red bar shows the predicted miR-18a binding sites in the 3’UTR of ATM. The sequence of mature miR-18a aligned to target sites is shown. (II) Luciferase activity assay. The reporter constructs in which the 3’UTR of ATM, wild-type or miR-18a binding-site mutant, was cloned downstream of the luciferase open reading frame. 293 cells were co-transfected with the luciferase construct and miR-18a mimics or control miRNA. The renilla construct was also cotransfected as an internal control. Luciferase activity was normalized to renilla luciferase activity. The data were presented as the means±SD of two experiments with six replicates (Student T-test, *p < 0.05; **p < 0.01; ***p < 0.001). III: miR-18a decreased the expression of ATM. Western blot of ATM was performed 48 h after the transfection of miR-18a mimics and miR-18a inhibitor. GAPDH was used as an internal control. c ATM inhibited the expression of EBV-related genes under hypoxic conditions (*p < 0.05; **p < 0.01). d ATM reversed the promotion effect of miR-18a on the EBV-related gene expression (*p < 0.05; **p < 0.01). e EBV gene expression after transfection of ATM under normoxia (*p < 0.05; **p < 0.01). f Transfection with ATM inhibited the promotion effect of miR-18a on the EBV viral load