Figure 5. Viral RNA and immediate early proteins associate with B5 in RNA-protein complexes.

(A) Cells were infected with HSV-2(G) and 3, 5, or 7 h pi, the cells were treated with 0.04% of methanol-free formaldehyde to cross-link RNA-protein complexes. The complexes were immunoprecipitated with a polyclonal rabbit anti-B5. and RNA was extracted from the supernatant and pellet and analyzed for the expression of viral transcripts and RPLPO. Results are shown for the time pi at which peak expression of viral transcripts was detected (3 h for ICP4, 5 h for ICP0 and 7 h for gB) and are presented as the ratio of viral transcript/RPLPO in each fraction to the viral transcipts/RPLPO in the total RNA-protein complex from infected cells prior to immunoprecipitation (input). Expression of RPLPO is presented as the ratio of RPLPO in each fraction to RPLPO in the total RNA-protein complex from uninfected cells prior to immunoprecipitation (input). (B) The supernatant and pellet from the immunoprecipitated complexes isolated at 0, 3, 5, and 7 h pi were evaluated for the expression of ICP4 (HSV-2), gB (HSV-2), ICP0 (HSV-1) or RPLPO by Western blots. Blots are representative of results obtained in at least 3 independent experiments.