Figure 6

The combined effect of G3 and Pam3CSK4 on IFN-γ production by eqPBMC when the two inducers were added simultaneously or sequentially. A The IFN-γ gene expression (FC values) obtained for sequentially stimulated cells were for each horse transformed to percentages of that recorded at simultaneous addition of G3 and Pam3CSK4. In the figure, FC values of simultaneously induced eqPBMC are displayed as 100%. Closed symbols represent cultures where G3 was added as first inducer (0 h) and Pam3CSK4 as second inducer (6 or 12 h). Open symbols represent cultures where Pam3CSK4 was added as first inducer (0 h) and G3 as second inducer (6 or 12 h). Total culture time was 18 h. Results are given as mean values ± SD, n = 6. B Protein concentrations of IFN-γ (ng/mL) in supernatants from eqPBMC cultured in the presence of G3 and Pam3CSK4 added simultaneously (0 h + 0 h), sequentially (0 h + 2, 4, 6 or 8 h) or separately (0, 2, 4, 6 or 8 h). Inverted pyramids = Pam3CSK4 alone; circles = G3 alone; squares = G3 followed by Pam3CSK4; pyramids = Pam3CSK4 followed by G3. For statistical analysis, the IFN-γ concentrations recorded in cultures exposed to Pam3CSK4 or G3 alone were pooled and compared with pooled values from cultures stimulated by the combinations of G3 and Pam3CSK4. Total culture time was 48 h. Results are given as mean values ± SD, n = 6. *P < 0.05 and ***P < 0.001.