Relative gene expression (A, C) and protein levels (B, D) of IFN-γ in eqPBMC cultured for 18 or 48 h in the presence of G3, Pam3CSK4, FliC and combinations thereof. The cytokine gene expression was normalized to the geometric mean for the reference genes (SDHA and RPL32) and calibrated to that in the medium control. The protein expression data are presented as the estimated IFN-γ levels after subtraction of the spontaneous IFN-γ production in parallel control cultures. Experiments for gene expression and protein analyses were conducted independently. Closed and open symbols indicate 3 µg G3/mL and 5 µg G3/mL, respectively. *P < 0.05.