Figure 7.

Neospora caninum extracellular vesicles (EVs) induce cytokine secretion in bone marrow-derived macrophages (BMDMs) by activating toll-like receptor 2 (TLR2) signaling pathway. (A) RT-PCR analysis of the relative mRNA level of TLR2 in total RNA isolated from 3 × 10⁶ BMDMs incubated with PBS, 10 µg/ml Pam3CSK4 or 50 µg/ml N. caninum EVs. The mRNA levels were normalized to those of GAPDH. (B) TLR2^−/− and wild-type (WT) mouse BMDMs were incubated with 50 µg/ml N. caninum EVs for 8, 12, and 24 h, and the supernatants were removed and assayed for IL-12p40, TNF-α, IL-1β, IL-6, IFN-γ, and IL-10 using ELISA assays. The data are representative of three independent experiments and are presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 for the TLR2^−/− group versus the WT group.