Fig. 1

ECTV CrmD recombinant viruses. a Schematic diagram of the genomic structure of the recombinant ECTVs generated for the study of the role of ECTV CrmD in mousepox pathogenesis. The names of the genes flanking the CrmD locus are indicated. In gray, the CrmD locus in the parental and recombinant viruses is shown. Both left inverted terminal repeat (lITR) and right inverted terminal repeat (rITR) are represented for ECTV, whereas the lITR is shown for the other viruses. b Western blot analyses using anti-CrmD and anti-35-kDa vCKBP antisera of supernatants from BSC-1 cells that were mock-infected (1) or infected with ECTV (2), ECTVΔCrmD (3), ECTVRevCrmD (4) or ECTVRevCRD (5) at a multiplicity of infection of 5 PFU/cell and harvested at 24 h post-infection. The position of the respective proteins is indicated by arrows. Molecular size markers in kDa are shown on the left. c TNF-induced cytotoxicity assay. Increasing amounts of recombinant ECTV CrmD (rCrmD, in μg/ml) or supernatants (equivalent to 5, 10 or 30 × 10³ cells) obtained as in b were added to block the effect of TNF on L929 cells. The values obtained in the presence of mock-infected cell supernatants have been substracted in each case. Data are mean +/− standard error of the mean (SEM) of triplicate samples. d Single-step growth curves of the indicated viruses. BSC-1 cells were infected with 5 PFU/cell and the virus production was titrated at the indicated hour post-infection. The means of duplicate samples are shown