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. 2020 Aug 16;7(19):2001402. doi: 10.1002/advs.202001402

Figure 4.

Figure 4

Effect of site‐directed removal of m6A modification on Virus replication. A) Measurement of m6A level on HBV pgRNA in individual groups. Relative ligation products of each group were quantified by qPCR. Bars represent mean ± SD from three independent experiments (n = 3, two‐sided Student's t‐test). B) cccDNA copy numbers of HBV in individual groups were measured by micro‐drop digital PCR analysis. In this part, cells were all transfected with GCN4‐dCas13b. scFv‐GGS6‐FTO (WT), scFv‐GGS6‐FTOH231A D233A (Mut), sgRNA (sg‐HBV), or pc0043 vector (sg‐Control) were transfected as indicated for overall 48 h. Bars represent mean ± SD from three independent experiments (n = 3, two‐sided Student's t‐test). C) Western‐blot results of HIV‐1 HXB2 P24 intracellular protein expression. Cells were all transfected as indicated for overall 48 h. This experiment was repeated three times with similar results. D) Measurement of m6A level on targeted HIV m6A site in individual groups. Relative ligation products of each group were quantified by qPCR. Cells were all transfected with GCN4‐dCas13b. scFv‐GGS6‐ALKBH5 (WT), scFv‐GGS6‐ALKBH5H204A(Mut), sgRNA (sg‐HIV), or pc0043 vector (sg‐Control) were transfected as indicated for overall 48 h. Bars represent mean ± SD from three independent experiments (n = 3, two‐sided Student's t‐test).