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. 2017 Apr 21;21(3):345–352. doi: 10.4196/kjpp.2017.21.3.345

Fig. 3. The suppressive effect of JS-III-49 on the NF-κB pathway is mediated by direct targeting of Akt.

Fig. 3

(A) RAW264.7 cells pretreated with JS-III-49 (30 µM) for 30 min were treated with LPS (1 µg/ml) for the indicated time, and protein levels of p65 and p50 in the nuclear fraction of the cells were determined by Western blot analysis. (B) RAW264.7 cells pretreated with JS-III-49 (30 µM) for 30 min were treated with LPS (1 µg/ml) for the indicated time, and phosphorylated and total levels of IκBα in the total cell lysate were determined by Western blot analysis. (C) RAW264.7 cells pretreated with JS-III-49 (30 µM) for 30 min were treated with LPS (1 µg/ml) for the indicated time, and phosphorylated and total levels of Src, Syk, and Akt in the total cell lysate were determined by Western blot analysis. (D) Effects of JS-III-49 on the kinase activities of Src and Syk were determined by a conventional in vitro kinase assay using purified Src and Syk as described in the Materials and Methods. (E) RAW264.7 cells pretreated with the indicated doses of JS-III-49 for 30 min were treated with LPS (1 µg/ml) for 5 min, and phosphorylated and total levels of Akt in the total cell lysate were determined by Western blot analysis. Lamin A/C and β-actin were used as internal controls for the nuclear fraction and total cell lysate, respectively.