Figure 7.

Anti-HBx gene delivery via transposons suppresses MALAT1-m6A-initiated HBV-related hepatocarcinogenesis in vivo. HBx-Tg mice were injected with 0.3 mL of TransIT-EE containing PB-20F3 (10 μg) and PB-Trans (5 μg) once every 4 days for a total of 7 injections. A Scheme of the plasmids PB-20F3, with SPSS used as a negative control. B Schematic diagram of the experimental design for HBx-targeting intervention in HBx-Tg mice. C Representative images showing fluorescence of mRuby3 in livers from the different groups, with nuclei counterstained with DAPI (blue). Scale bars: 100 μm. D The relative expressions of MALAT1 in the livers was detected by qRT-PCR. E Representative images showing FISH assays of the RNA levels of MALAT1 (red) in the livers, with nuclei counterstained with DAPI (blue). Scale bars: 100 μm. F The overall m6A content in the livers was analyzed by m6A quantitation analysis. G MeRIP assays were performed in the livers, while the abundance of MALAT1 with anti-m6A antibodies was measured by qRT-PCR and normalized to that of IgG. H RIP assays were performed with IGF2BP3 antibody in the livers, while the abundance of MALAT1 was measured by qRT-PCR. I The levels of HBx, METTL3, METTL14, WTAP, IGF2BP3, E-cadherin, and Vimentin proteins in the livers were detected by WB. J Representative images showing HBx, METTL3, METTL14, WTAP, IGF2BP3, E-cadherin, and Vimentin immunostaining of livers. Scale bars: 100 μm. *P < 0.05; **P < 0.01; ***P < 0.001. n.s., not significant.