Fig. 3
From: Replication of cowpox virus in macrophages is dependent on the host range factor p28/N1R
CPXV replication in primary macrophages. The replication of a CPXV-∆p28, CPXV-p28(1-184) in primary peritoneal rat macrophages and b of CPXV-∆p28 in human PBMC-derived GM-CSF-differentiated macrophages was analysed via quantification of viral genome replication in cell lysates (n = 2). Primary rat macrophages were isolated from the peritoneum of 3-month-old Wistar rats (Rattus norvegicus). PBMCs were isolated from fresh human buffy coat (acquired from DRK Blutspendedienst Ost) via Ficoll-Paque density gradient cell separation. Isolation of CD14+ monocytes was done via MACS separation using CD14+ MicroBeads (Miltenyi Biotec). For differentiation into macrophages, 100 ng/ml GM-CSF was added to the medium. Cells were infected with an MoI of 0.1 and viral genome copies in cell lysates were quantified via qPCR. Shown are fold-change values obtained via the ∆∆cT method after normalization of cT values to MYC gene expression. Statistics: a one-way ANOVA and Tukey’s multiple comparison test, b unpaired t-test