Viral mRNA and protein expression in H6R28LEP-infected PBMCs.

<p><b>A:</b> PBMCs and CMBCs were infected with H6R28LEP, total RNA was purified after 24 h, and mRNA expression of immediate early (U90, U94), early (U41, U69), and late (U24, U39, U48) genes was assayed by real-time PCR after RT. The data are shown as mRNA expression volumes in PBMCs relative to those in CBMCs. Data are given as mean ±1 standard deviation (<i>n</i> = 3). <b>B:</b> At 24 h after infection, H6R28LEP-infected CBMCs or PBMCs were fixed in cold acetone and expression of the viral proteins U90, U39, U41, and U48 was observed using an indirect immunofluorescent assay. <b>C:</b> Proportion of H6R28LEP intact virus particles produced in the culture supernatant. CBMCs or PBMCs were infected with H6R28LEP, and real-time PCR was used to quantify the number of H6R28LEP genome DNA copies present in the supernatant after 3 d. The supernatant from each culture was used to infect PBMCs by using centrifugation (left) or the normal adsorption method (right), and the H6R28LEP intact virus particles in the supernatant were assayed. The graph shows the percentage of intact virus particles relative to H6R28LEP genome DNA copy numbers in the supernatant. Data are given as mean ±1 standard deviation (<i>n</i> = 3).