Fig 2. Detection of the full length L protein in NSDV-infected cells.

Vero cells were infected with the NSDVi isolate at a MOI of 6 TCID₅₀ or left uninfected (uninf.). After 16 h, infected and uninfected cells were harvested using SDS Sample Buffer containing protease inhibitors, and proteins were separated on 5% acrylamide SDS-PAGE gels and transferred onto polyvinylidene difluoride (PVDF) membrane. The membrane was cut vertically along the middle of the track containing proteins from infected cell lysate. Blots were then incubated with the indicated antiserum or purified antibodies before developing with HRP-anti-rabbit IgG. (A) Filters were incubated with diluted antiserum raised against the N-terminus of the L protein (anti-L(NT)) or the C-terminus of the L protein (anti-L(CT)). (B) Filters were incubated with the pre-immune sera corresponding to the sera used in A. (C) Membranes were incubated with affinity purified antibodies extracted from the sera used in A. For all immunoblots the migration position of protein size markers are indicated. The star (*) indicates a non-L protein peptide labelled by crude antiserum but not by affinity-purified antibody.