MDCKII cells (H1N1v):74.7 ± 1.0 μM (EC50)|MEK2:60 nM (cell free)|MEK1:70 nM (cell free)|A549 cells (H1N1v):1.2 ± 0.4 μM (EC50)

方法：为研究U0126-EtOH的抗肿瘤活性，每天将U0126-EtOH（10.5 mg/kg）腹腔注射给小鼠治疗。 结果：U0126-EtOH导致肿瘤植入和早期生长显著减少，注射9天后的肿瘤体积减少了60-70%，此后一直保持这种状态。[4] 方法：为研究U0126-EtOH对血管收缩的影响，大鼠接受120分钟的暂时性中脑动脉阻塞（tMCAO），再将U0126-EtOH（30 mg/kg）腹腔注射给大鼠。 结果：用U0126-EtOH处理后，对S6c的血管收缩显著减少。[5]

方法：COS-7细胞用U0126-EtOH处理后检测AP-1转录活性。 结果：U0126-EtOH抑制AP-1转录活性（IC50=1 μM）。[1] 方法：HCT116细胞用U0126-EtOH处理后使用软琼脂生长实验检测克隆形成，HeLa细胞用U0126-EtOH处理后检测Elk1-荧光素酶报告基因。 结果：U0126-EtOH抑制贴壁非依赖性集落形成（IC50=19.4 μM），U0126-EtOH抑制EGF刺激的Elk1荧光素酶报告基因（IC50=0.29 μM）。[2] 方法：小鼠RAS-3T3细胞用U0126-EtOH处理后使用ELISA方法检测ERK1/2磷酸化水平。 结果：10-40 μM的U0126-EtOH抑制MEK介导的ERK1/2磷酸化。[3]

DMSO:255 mg/mL (597.75 mM);10% DMSO+40% PEG300+5% Tween 80+45% Saline:7.9 mg/mL (18.52 mM);Ethanol:< 1 mg/mL (insoluble or slightly soluble)

HEK293 cells were maintained in Dulbecco's modification of Eagle's medium (low glucose) plus 10% foetal bovine serum. HeLa cells stably expressing wild type or kinase-dead LKB1 have been described. AMPK activity was determined by immunoprecipitate kinase assays using anti-AMPK-a1 and -a2 antibodies. Antibodies recognising AMPK phosphorylated on Thr-172 (anti-pT172), AMPK-α1 and -α2 and acetyl-CoA carboxylase-1 (ACC1) phosphorylated on Ser-80 [16] were described previously. Quantification of ratios of signals from phosphorylated and total protein using these antibodies was performed by dual labelling using the LI-COR Odyssey IR imager as described. Contents of ATP and ADP were determined for cells in 6 cm culture dishes by quickly pouring off the medium, adding 350 μl of ice-cold 5% perchloric acid, scraping the cells off with a plastic scraper, and centrifuging (14 000 · g; 3 min, 4 °C) to remove insoluble material. The perchloric acid was then extracted from the supernatant and nucleotides analysed by capillary electrophoresis of perchloric acid extracts as described previously. All incubations of cells were performed in triplicate and results are expressed as means ± S.E.M [3].

Prior to injection, FI cells were labeled with a stable fluorescent dye molecule, DiA at 10 μg/ml for 5 h at 37 1C. After washing to remove free DiA, cells were trypsinized for inoculation (U0126 experiments) or transfection (RNAi experiments). Biliary epithelial cells were injected subcutaneously, at the indicated times, into the tibia of nude mice. In the chemical experiments, 3h after inoculation, mice were treated with U0126 (10.5 mg/kg) daily by intraperitoneal injection. The length and width of each tumor were measured every day by using a caliper. The following formula was used to calculate tumor volumes ? width2 length/2. Mice were killed at the end of experiment. Tumors were immediately frozen in liquid nitrogen [5].