Figure 3. Effect of macrophage activation stimulated with water-insoluble -glucans on suppression of TLR2, TLR4, NOD1, and NOD2 mRNA expression. (A) The primers used in the RT-PCR assay and sequence of the target mRNA for TLR2, TLR4, NOD1, and NOD 2. (B) J774.1 cells were transfected with siRNA of TLR2, TLR4, NOD1, NOD2, or buffer only with the use of TransIT-TKO transfection reagent. Total RNA was extracted 24 hrs after transfection, and RT-PCR was performed. (C) J774.1 or transfected J774.1 cells (10⁵ cells/mL) were stimulated with 1 mg/mL of water-insoluble -glucan for 24 hrs, and the TNF- in the culture supernatants was measured by a TNF- ELISA kit.