Figure 3

Internalisation of HEV-VLPs in different cell lines. a Confocal microscopy images for checking the expression of RFP in RNA-VLP [5′-methyl-G-HEV(1–249 nt)-RFP RNA and pORF2 VLP’s] incubated i Huh7, ii A549, iii HeLa, iv Vero and v SiHa cells. b Confocal microscopy images for RFP fluorescence in Huh7 cells incubated with i 5′-methyl-G-HEV (1–249 nt)-RFP RNA only; RNA-VLP complexes prepared from ii 5′-methyl-G-HEV (199–444 nt)-RFP RNA and pORF2-VLPs, iii 5′-methyl-G-HEV (1–249 nt)-RFP RNA and Δ112aa pORF2-VLPs, iv 5′-methyl-G-HEV(1–249 nt)-RFP RNA, annealed with complementary oligo (HEV, 150–200 nt) and pORF2-VLPs. c Fluorescent microscopy images (i–iii) of Immunofluorescence assay for HBV surface antigen (HBsAg) in Huh 7 cells, incubated with HEV(1–249 nt)-HBsAg-RNA-pORF2 VLPs. Indirect immunofluorescence assay was performed for detecting HBsAg expression using anti-HBsAg primary antibody followed by Alexa-546 conjugate secondary antibody.