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

Figure 6

Effects of ISE on the activation of NF-κB p65 and MAPK in RAW264.7 cells. (a, b) Protein expression of p-p65 and p65 was analyzed through western blotting. HDAC1 and GAPDH was used as an internal control. (c) Results of the NF-κB reporter assay of ISE- and LPS-treated cells grown in 96-well plates, which were cotransfected with a pNF-κB-Luc vector and a pNL-Luc vector using the FuGENE 6 Transfection Reagent. After cotransfection for 24 h, cells were treated with growth medium containing the indicated concentration of ISE or 10 ng/mL of LPS 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. (d) Protein expression of p-ERK1/2, ERK1/2, p-p38, p38, p-JNK, and JNK was analyzed through western blotting. GAPDH was used as an internal control. Data are shown as of triplicate experiments.

(a)

(b)

(c)

(d)