Transient Reversal of Episome Silencing Precedes VP16-Dependent Transcription during Reactivation of Latent HSV-1 in Neurons
Figure 2
During reactivation, HSV-1 exhibits a biphasic profile of viral transcripts in SCG neurons.
(A) Scheme showing a typical reactivation experiment. Neuron cultures were established and then infected with HSV-1 GFP-Us11 (MOI = 1) in the presence of 100 µM acyclovir (ACV). Latency was established over a 7-day period before re-feeding with fresh media lacking ACV. The next day, reactivation was induced with 20 µM LY294002. (B) Profile of viral mRNA accumulation in response to LY294002. RNA was collected at the indicated times and analyzed by qRT-PCR. Values are normalized against the 0 h sample [ICP27, 171 copies/sample; UL5, 135 copies/sample; UL30, 94 copies/sample; VP16, 347 copies/sample and UL36 130 copies/sample]. Data is derived from three or more independent cultures and reactivation experiments. (C) Reactivation profiling in the presence of the viral DNA encapsidation inhibitor WAY150138 (20 µg/ml). (D) Transcript levels at 20 h post induction in the absence (−) or presence (+) of protein synthesis inhibitor cyclohexamide (CHX, 10 µg/ml). To ensure cell viability, CHX was added 10 h after LY294002, prior to the appearance of new viral transcripts.