Figure 3
NiV proteins are released in the absence of MVA. 293T cells were transfected with pCAGGS constructs containing NiV genes. (A) Protein released into the supernatant was purified by gradient centrifugation as described in Methods. Proteins were immunoprecipitated with MAbs F45G6 (anti-N), F45G5 (anti-M) or rabbit polyclonal anti-serum against NiV F or HeV G. (B) Protein released as in (A) was quantitated by densitometry. The mean and standard deviation of three or more experiments are shown. ND = not detectable. Release of protein from cells expressing combinations of F and G (C) N, M, F, and G (D) or N and M (E) are shown along with the percent protein released. N was not immunoprecipitated in D to allow visualization of F0. Quantitation for C was performed on a lighter exposure. Proteins derived from cell lysates (L) or culture supernatants (S) are indicated. Lysate bands represent 1/6 of total lysate. Representative experiments are shown.