Figure 6

Role of TNF-α in inducing CXCL10 expression. (a) MuV-induced TNF-α and CXCL10 production. Sertoli and germ cells of 4-week-old WT or TNF-α^−/− mice were co-cultured and infected with MuV or with MuV in the presence of pomalidomide (POM), an inhibitor of TNF-α secretion for 24 h. Co-cultures without MuV infection served as controls (Ctrl). TNF-α (left panel) and CXCL10 (right panel) levels in the culture media were measured using ELISA. (b) MuV-induced germ cell apoptosis. Co-cultures were treated as described in a. Apoptotic germ cells were determined based on AO/EB staining and confirmed by determination of caspase cleavages by Western blot analysis. (c) Induction CXCL10 production by recombinant TNF-α. WT or TNF-α^−/− cells were co-cultured in the presence of the indicated doses of recombinant mouse TNF-α for 24 h. CXCL10 levels in media were determined using ELISA. (d) TNF-α-induced germ cells apoptosis. WT or TNF-α^−/− cell co-cultures were treated with 5 ng/ml TNF-α or with TNF-α in the presence of Ab-CXCR3 for 24 h. Apoptotic germ cells were determined based on AO/EB staining and caspase activation was assessed by western blot analysis. Data are the means±S.E.M. of three independent experiments. *P<0.05 and **P<0.01