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. 2020 Aug 25;16(8):e1008778. doi: 10.1371/journal.ppat.1008778

Fig 5. MT1G and other MT1 genes suppress lytic replication of EBV.

Fig 5

(A) Protein expression of V5-tagged MT1G in stably transduced AGS-BX cells was verified by immunoblotting. (B) Newly produced EBV viruses from TPA/NaB treated or untreated AGS-BX cells stably expressing V5-MT1G or the vector control were titrated using Raji cells via flow cytometry. (C) Representative diagram of flow cytometry for (B). AGS cells were used to determine the background. (D) EBV genome copies (EBNA1 DNAs) in TPA/NaB treated AGS-BX cells stably expressing V5-MT1G or the vector control in the presence of Zinc (50uM) were measured by qPCR assays at 24 hpi (left panel) and 48 hpi (right panel). The relative level of EBNA1 DNAs in TPA/NaB treated AGS-BX cells transduced with the vector control was set up as 1. (E) Expression of EBV lytic genes (BZLF1, BRLF1, BMRF1, BALF5, BNRF1, BcLF1, BLLF1) in TPA/NaB treated or untreated AGS-BX cells stably expressing V5-MT1G or the vector control in the presence of Zinc (50uM) were measured by RT-qPCR assays at 24hpi. (F) Protein expression of V5-tagged MT1 genes (MT1F, H, M, X) in stably transduced AGS-BX cells was verified by immunoblotting. (G) EBV genome copies in TPA/NaB treated AGS-BX cells stably expressing V5-tagged MT1 genes (MT1F, H, M, X) or the vector control in the presence of Zinc (50uM) were measured by qPCR assays at 24hpi, similar as (D). (H) Expression of EBV lytic genes in TPA/NaB treated or untreated AGS-BX cells stably expressing V5-tagged MT1 genes (MT1F, H, M, X) or the vector control in the presence of Zinc (50uM) were measured by RT-qPCR assays at 24hpi, similar as (E). All of above results are based on n = 3 independent repeats and presented as mean ± SEM (* p<0.05; ** p<0.01; *** p<0.001, two-tailed paired Student t-test).