Figure 3

Vaccinia virus-induced HSE binding activity in macrophages and a human hepatoma cell line. (A) Macrophages were infected with vaccinia virus (V) with 5 pfu/cell and cultured for 16 or 24 hours (supershift assay). Lane 1 – NE from infected macrophages at 16 h p.i., lane 2 – WCE from control macrophages, lane 3 – WCE from infected macrophages at 16 h p.i., lane 4 – WCE from infected macrophages at 24 h p.i.; lanes 5–8 – as lanes 1–4 plus preincubation with 1:20 dilution of antibodies against HSF1 (aHSF1). (B) HepG2 cells (3 × 10⁶) were infected with vaccinia virus at MOI 1 for 24 h or heat shock treated (44°C, 20 min) or heat shock treated and recovered for 2 or 4 h at 37°C (shift assay). Lane 1 – control cells, lane 2 – heat shock treated, lane 3 – heat shock treated and recovered for 2 h, lane 4 – heat shock treated and recovered for 4 h, lane 5 – vaccinia virus infected for 24 h. O – ds oligoDNA (free HSE) incubated without proteins. Exposure time: 6 days (A) and 18 h (B). A single representative experiment being one of four separate experiments is shown.