FIG. 2.

Transcription and translation of RSV G and F glycoprotein genes in PIV-RSV subgroup A chimeric viruses and in RSV. Accumulation of RSV G and F mRNAs (A) and proteins (B and C) was examined in HEp-2 cells infected with rB/HPIV3 (lanes 1), rB/HPIV3-G_A (lanes 2), rB/HPIV3-F_A (lanes 3), rB/HPIV3-G_AF_A (lanes 4), and RSV (lanes 5) at an MOI of 5, and cells were harvested at the indicated time points postinfection. mRNAs were detected with a mixture of RSV G and F specific ³²P-labeled DNA probes (A). The F mRNA transcribed from rB/HPIV3-F_A and rB/HPIV3-G_AF_A is shorter than that transcribed from RSV because the 5′ nontranslated region of F was deleted in the chimeric viruses. G and F glycoproteins were detected by a mixture of antisera raised against the cytoplasmic domain of RSV A2 F protein and against the ectodomain of the RSV A2 G protein (B). Glycoproteins were visualized by incubation with horseradish peroxidase-labeled goat anti-rabbit IgG antibodies. Since the F₁ subunit of the fusion glycoprotein and the partially glycosylated G glycoprotein have similar electrophoretic mobilities, an additional Western blot was stained with F-specific antiserum only (C).