FIG. 4.
FSK's action is immediate and does not require de novo protein synthesis. Time course for FSK-induced expression of MIE RNA (A) and protein (B). FSK (10 μM) was added to or omitted from (No Add) quiescently infected NT2 at 24 h p.i. RNA and protein were isolated at the designated times thereafter. MIE RNA was quantified by real-time RT-PCR, using the standard-curve method and normalization to the 18S RNA amount (RNAN). The IE1 p71 and IE2 p86 proteins were analyzed by the immunoblotting method specified in the legend of Fig. 3C. MK, mock-infected NT2. (C) FSK-induced MIE RNA expression in the absence of protein synthesis. FSK (10 μM), CH (200 μg/ml), or both FSK and CH were added to or omitted from (No Add) quiescently infected NT2 at 24 h p.i., except that CH was added 30 min prior to the addition of FSK. MIE RNA was quantified at 4 h after FSK addition, using methods described above for panel A. Error bars represent standard errors of the means from two experiments.