FIG 5.

Effect of LNA-miR-BART20-5pi on BZLF1 and BRLF1 expression. (A) miR-BART20-5p expression before and after lytic induction. To induce the lytic cycle, AGS-EBV cells were treated with 5 nM TPA for 48 h. (B, C) Effect of LNA-miR-BART20-5pi on the level of miR-BART20-5p (B) and the BZLF1 and BRLF1 mRNAs (C). AGS-EBV cells were transfected with 30 nM LNA-miR-BART20-5pi. At 24 h after transfection, the cells were treated with 5 nM TPA. Real-time RT-PCR was carried out 48 h later to assess the level of miR-BART20-5p (B) or the BZLF1 and BRLF1 mRNAs (C). (D) Effect of LNA-miR-BART20-5pi on expression of various EBV proteins. The same sets of cells used for the assays whose results are presented in panels B and C were used for Western blotting. The numbers under each band represent the ratio of the level of expression to that for cells transfected with the control inhibitor. (E) Experiments similar to those described for panel C were also performed using naturally EBV-infected gastric carcinoma cell lines SNU-719 and YCCEL1. For SNU-719 and YCCEL1 cells, 50 nM LNA-miR-BART20-5pi was transfected. Error bars indicate SDs. *, P < 0.05; †, P < 0.01.