Figure 6.

Effect of silencing the expression of 3‐OST2, HVEM and nectin‐2 on HSV‐1 gD‐ and CyPB‐induced signalling in macrophages. The contribution of 3‐OST2 (as a 3‐O‐sulfated HS‐generating enzyme) and of HVEM and nectin‐2 to the activation of signalling pathways was evaluated by analysing the phosphorylation of Akt and ERK1/2 in siRNA‐transfected macrophages. Following treatment with siRNA for 48 h, cells were stimulated with HSV‐1 gD or CyPB (both at 1 μg·mL⁻¹) for various times, and the phosphorylation of ERK1/2 (P‐ERK1/2) and Akt (P‐Akt) was analysed by western blot. Parallel immunoblotting with antibodies to Akt and ERK1/2 confirmed equal loading of samples. Data are representative of three independent experiments. The ratio of P‐Akt/Akt and P‐ERK1/2/ERK1/2 were quantified and normalized to unstimulated cells.