FIG. 5.

Viral RNA and protein expression over time during infection with WT and mutant HPIV1. A549 cells were infected with WT, F170S, or P(C−) HPIV1 at an MOI of 5 for 12, 24, or 48 h. Relative amounts of HPIV1 (A) combined genomic and antigenomic RNA, (B) N mRNA, and (C) P mRNA were measured by real-time PCR. The values were normalized to the 12-h time point for WT HPIV1, which was assigned a value of 1.0. (D) HPIV1 N, C, and P proteins were detected by Western blotting from samples collected in parallel to the samples in panels A to C. Shown is a representative of two independent experiments with similar results. (E) A549 cells were infected with F170S HPIV1 at an MOI of 5 for 24, 30, 36, 42, or 48 h. The C and P proteins were detected by Western blotting. P denotes the position of the P protein, n.s. indicates a nonspecific band, and the asterisk indicates a potential P cleavage or degradation product detected by the polyclonal anti-P antibody.