In vitro multiplication and sFL production by rVACV. Confluent CV-1cell cultures were infected with purified virus at a MOI of2.5 at 37°C for 1 hour, washed with PBS and the fresh medium was added. The medium and cells were collected, frozen and thawed, and cell debris was removed. The titers of the infectious virus (A) were determined in the medium and in the cell lysate. Total FL production was determined by ELISA (B). The columns in all graphs represent the mean ± s.d. The intracellular location of sFL (C) in 293T cells (a, b) 48 h after transfection or in infected CV1 (c, d) and HeLa cells (e, f) 9 h after infection or in J774.G8 cells (g, h) 3 h after infection with P13-H5-FL (c, e, g) or P13-E/L-FL (d, f, h) as visualized by an immunofluorescent microscope at a magnification of 1000×. The colocalisation of sFL (D) with endoplasmic reticulum marker calreticulin or with cis-Golgi marker GM130 in HeLa cells 3 h after infection as visualized by an immunofluorescent microscope at a magnification of 1000×.
Zurkova et al. Virology Journal 2010 7:109 doi:10.1186/1743-422X-7-109