IL-12p35 is a TLR3/TLR4 primary response gene. Monocyte-derived DCs were either incubated in medium alone or stimulated with LPS (1 μg/mL), PGN (10 μg/mL), Pam₃CSK₄ (10 μg/mL), or PolyI:C (10 μg/mL). (A) Total RNA was extracted after 3 and 6 hours and analyzed by real-time RT-PCR. Levels were normalized using β-actin mRNA as reference and compared with unstimulated conditions. Data are shown as mean ± SEM of 6 independent experiments on different donors. (B) DCs were pretreated with CHX (10 μg/mL) for 1 hour before LPS or PolyI:C addition for 3 hours. Total RNA was then extracted and analyzed by real-time RT-PCR. One representative donor of 5 is shown.