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. Author manuscript; available in PMC: 2015 Jan 20.
Published in final edited form as: Virology. 2013 Dec 20;449:304–316. doi: 10.1016/j.virol.2013.10.038

Figure 1. 293TTF cell line overexpresses furin and facilitates production of furin-cleaved PsV.

Figure 1

Expression of endogenous furin in parental 293TT cells (lane 1), 293TT transiently transfected with furin gene (lane 2), 293TTF cells stably transfected with furin in the presence (lane 3) and absence (lane 4) of puromycin (2μg/ml). Equivalent amounts of conditioned media supernatant of parental 293TT (lane 5) or 293TTF in the presence (lane 6) and absence (lane 7) of puromycin (2μg/ml) were used to assess furin secretion. Image analyses and relative densitometry using ImageJ indicate that for 293TTF cells endogenous furin expression and secretion was at least 150-fold higher compared to 293TT control cells. Band seen at ~60kDa was a furin splice variant as indicated in the material data sheet of the antibody used (A). Equivalent amounts of HPV16 PsV (solid squares) or HPV16 fcPsV (open squares) based on L1 content, were added onto pre-plated furin deficient cells FD11 (B) or FD11F (FD11 cells re-complemented with furin gene) (C), 293TT cells (D) and 293TT cells with 20μM of furin inhibitor (E). All experiments were performed in triplicate