0.01

1. DBM1285 [cyclopropyl-{4-[4-(4-fluorophenyl)-2-piperidin-4-yl-thiazol-5-yl]pyrimidin-2-yl}amine] suppresses tumor necrosis factor-a production by blocking p38 mitogen-activated protein kinase/mitogen-activated protein kinase-activated protein kinase 2 signaling pathway, Kang JS, Kim HM, Choi IY, Han SB, Yoon YD, Lee H, Park KH, Cho IG, Lee CW, Lee K, Lee KH and Park SK, Journal of Pharmacology and Experimental Therapeutics Supplementary Fig. 2 A B LPS (1 mg/ml) + DBM1285 (mM) LPS (1 mg/ml) + DBM1285 (mM) UN VH 0.01 0.1 1 UN VH 0.001 0.01 0.1 1 TNF-a p-p38 MAPK b-actin Effect of DBM1285 on LPS-induced TNF-a mRNA expression and p38 MAPK phosphorylation in RAW 264.7 cells. RAW 264.7 cells were plated at 5 X 105 cells/ml. Vehicle (VH, DMSO) or indicated concentrations of DBM1285 (0.001, 0.01, 0.1 or 1 mM) was pretreated for 1 h before being incubated with LPS (1 mg/ml) for 6 h or 30 min. (A) Total RNAs were isolated and the mRNA expression of TNF-a and b-actin was determined by RT-PCR. (B) Total cell lysates were prepared and phosphorylated forms of p38 MAPK was detected by Western immunoblot analysis.