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Fig. 1 | Virology Journal

Fig. 1

From: Human cytomegalovirus reactivation from latency: validation of a “switch” model in vitro

Fig. 1

Time-course changes in HCMV DNA nuclear amounts in THP-1 latency and reactivation models. a At the indicated time points p.i., undifferentiated (panel “THP-1 latency model”) and differentiated THP-1 cells (panel “THP-1 reactivation model”) were subjected to cellular fractionation in order to obtain purified nuclei (Nuc) and cytoplasmic (Cyt) fractions derived from total cell lysates (Lys). After SDS-PAGE, WB analysis was done using a rabbit polyclonal antibody directed to B23 (40 kDa) and a mouse monoclonal anti-beta-actin (44 kDa) antibody as nuclear and cytoplasmic markers, respectively. The immunoreactions were revealed by AP-conjugated anti-rabbit and anti-mouse antibodies. Molecular weight markers are indicated on the left-hand figure. b Total DNA was extracted from nuclei of undifferentiated THP-1 monocytes (panel “THP-1 latency model”) and THP-1 monocytes differentiated after infection (panel “THP-1 reactivation model”). HCMV DNA amounts were measured by q-Real-time PCR at 30 h, 4, 6 and 7 days p.i. at MOI 0.5, 0.25 and 0.125 in both experimental models. Comparable results were obtained from two independent experiments. c Total DNA was extracted from the nuclei of “viable” HCMV-infected THP-1 monocytes (“untreated virus”) and UV-inactivated HCMV-infected THP-1 monocytes (panel “THP-1 latency model, MOI 0.5”) and THP-1 monocytes differentiated after infection (panel “THP-1 reactivation model, MOI 0.5”). HCMV DNA amounts were measured by q-Real-time PCR at 30 h, 4, 6 and 7 days p.i. at MOI 0.25 in both experimental models

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