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. 2024 May 3;15:3729. doi: 10.1038/s41467-024-48031-8

Fig. 3. Reactivation of EBV lytic genes by LNP-encapsulated nucleoside-modified mRNA (mTZ3-LNP).

Fig. 3

a The particle size, polydispersity index, and zeta potential of mTZ3-LNP were analyzed using a dynamic light scattering method. The data are representative of four independent experiments. Data are presented as mean ± SD. b The cell uptake process of the LNP-encapsulated Cy5-labeled mTZ3 mRNAs by SNU719 cells over 1, 3 and 6 h was visualized using an LSM 880 confocal laser scanning microscope (n = 3 experimental replicates). The fluorescence signals were measured in three channels: Cy5, excitation/emission wavelength (ex/em), 633/697 nm; Dnd-26, ex/em, 488/524 nm; and hoechst, ex/em 405/460 nm. Scale bar = 10 μm. The signal of Cy5-labeled mTZ3 mRNAs in the cytoplasm of SNU719 cells is indicated by a white arrow. c Western blotting was used to detect Zta expression in SNU719 and C666-1 cells treated with mTZ3-LNPs and LNP-encapsulated control mRNA (control-LNP) for 24 h. The EBV-negative NPC cell line HK1 was used as the negative control (n = 3 experimental replicates). d Western blotting detected the expression of Zta, Rta, the downstream early (BGLF4) and late (VCA) lytic proteins and cleaved caspase 3 in SNU719 and C666-1 cells treated with mTZ3-LNPs for 3 to 96 h (n = 3 experimental replicates). e Immunofluorescence staining and (f) flow cytometry analysis illustrated the induction of Zta, EA-D, and gp350 expression in mTZ3-LNP-treated SNU719 and C666-1 cells (n = 3 experimental replicates). Representative IF images and flow cytometry plots are shown. Scale bar = 10 μm. Source data are provided as a Source Data file.