Figure 5

Characterisation of CVB3-VLPs produced by co-transfection of 3CD and P1. (A) SDS-PAGE and Western blot analyses of purified VLPs. The top panel shows the stain-free total protein staining of the purified CVB3-VLPs. The bottom panel shows VP0, VP1 and VP3 capsid protein detection by Western blot using an in-house produced rabbit anti-CVB1–6 polyclonal antibody. 2 µg of each purified CVB3-VLP was loaded per well recovered from cultures produced as follows (1) 2% 3CD/98% P1, 5 days production, (2) 2% 3CD/98% P1, 8 days production (3) 5% 3CD/95% P1, 5 days production, (4) 5% 3CD/95% P1, 8 days production (5) 10% 3CD/90% P1, 5 days production, 6) 10% 3CD/90% P1, 8 days production. The gel and blot were cropped for clarity. For full photos, see Supplementary Fig. 5. (B) Dynamic light scattering analysis of the purified CVB3-VLPs. Transmission electron microscopy images of the sucrose cushion ultracentrifugation purified CVB3-VLPs: (C) 2% 3CD/98% P1, 5 days production, (D) 2% 3CD/98% P1, 8 days production (E) 5% 3CD/95% P1, 5 days production, (F) 5% 3CD/95% P1, 8 days production (G) 10% 3CD/90% P1, 5 days production, (H) 10% 3CD/90% P1, 8 days production Scale bar 200 nm, 50,000 × magnification.