Anti-Inflammatory Effects of Ursolic Acid Figure 6. Inhibition of proliferation/survival associated signaling molecules by ursolic acid in activated T cells. Lymphocytes were incubated with UA (5 mM, 4 h) and stimulated with Con A (5 mg/ml) for 1 h. Whole cell lysates were prepared, fractionated on 10% SDS–PAGE, and electrotransferred to nitrocellulose membrane. Western blot analysis was performed using different antibodies specific for (A) p-c-Raf, p-MEK, p-ERK, ERK, p-JNK and JNK (B) IkBa and b-Actin (loading control). (C–E) Lymphocytes were incubated with 5 mM UA for 4 h and then stimulated with Con A (5 mg/ml) for 1 h. Nuclear extracts were prepared and analysed for NF-kB (B), NF-AT (C) and AP-1 (D) activation by EMSA. (E&F), UA inhibits NF-kB dependent gene products in activated lymphocytes. Lymphocytes were incubated with UA (5 mM, 4 h) and were stimulated with Con A (5 mg/ml) for PLoS ONE | www.plosone.org 10 February 2012 | Volume 7 | Issue 2 | e31318