Insampaedok-San Extract Exerts an Immune-Enhancing Effect through NF-κB p65 Pathway Activation

Figure 7

Effects of rapamycin on ISE-induced NF-κB transactivation and mRNA expression of cytokines in RAW264.7 cells. (a) RAW264.7 cells grown in 96-well plates were cotransfected with a pNF-κB-Luc vector and a pNL-Luc vector using FuGENE 6 Transfection Reagent. After cotransfection for 24 h, cells were treated with growth medium containing 400 μg/mL of ISE or 10 ng/mL of LPS with absence or presence of 50 μM of rapamycin for 6 h. Following LPS or ISE treatment, the cells were lysed and luciferase activity was assessed using a dual-luciferase reporter assay system and a luminometer. (b, c) mRNA expression of TNF-α and IL-6 was analyzed using qRT-PCR. GAPDH was used as an internal control. Data are shown as of triplicate experiments.

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