BAY 11-7082 (BAY 11-7821) is an NF-κB inhibitor that inhibits TNFα-induced IκBα phosphorylation (IC50=10 μM). BAY 11-7082 is also an inhibitor of the ubiquitin-specific proteases USP7 and USP21 (IC50=0.19/0.96 μM).

IκBα phosphorylation:10 μM

方法：人结直肠癌细胞 HT29 用 BAY 11-7082 (10-100 µM) 处理 1 h，随后用 TNF (50 ng/mL) 刺激 3-10 min 或用 Flag-TWEAK (200 ng/mL) 处理 8 h，使用 Western Blot 检测靶点蛋白表达水平。
结果：Bay 11-7082 抑制 TWEAK 诱导的 p100 加工和 TNF 诱导的 IκBα 的磷酸化和降解。在 Bay 11-7082 的浓度为 30-100 µM 时，两种 NFκB 途径均被显著阻断。[1]
方法：巨噬细胞 RAW264.7 用 BAY 11-7082 (15 µM) 和 LPS (1  µg/mL) 处理 6 h，使用 ELISA Assay 检测 TNF-α 水平。
结果：BAY 11-7082 阻断了 LPS 处理的 RAW264.7 细胞中 TNF-α 的产生，这是由活化的 NF-κB 产生的炎症反应。[2]

方法：为研究免疫调节作用，将 BAY 11-7082 (5-10 mg/kg) 腹腔注射给 C57BL/6 小鼠，1 h 后静脉注射 poly U (50 µg/head) + in vivo-jetPEI。
结果：当 TLR 配体刺激时，用 BAY 11-7082 处理可以通过限制 pDC 功能来抑制体内 IFN 反应。[3]
方法：为研究对银屑病样皮炎的作用，将 BAY 11-7082 (20 mg/kg) 腹腔注射给 IMQ 诱导银屑病样病变的 C57BL/6 和 NLRP3 KO 小鼠，每天一次，持续七天。
结果：BAY 11-7082 钝化表皮厚度、棘皮病和炎症浸润。BAY 11-7082 降低了 pNF-κB、NLRP3、TNF-α、IL-6 和 IL-1β 的表达，减弱了信号转导子和 STAT3 的磷酸化，并降低了 IL-23 水平。[4]

UBE1 (0.17 μM) in 22.5 μL of 20 mM Hepes, pH 7.5, containing 10 μM ubiquitin is incubated for 45 min at 21°C with 1 μL of DMSO or 1 μL of BAY 11-7082 in DMSO. A 2.5 μL solution of 10 mM magnesium acetate and 0.2 mM ATP is added, incubated for 10 min at 30°C, and the reactions are terminated by the addition of 2.5 μL of 10% (w/v) SDS and heating for 6 min at 75°C. The samples are subjected to SDS/PAGE in the absence of any thiol. The gels are stained for 1 h with Coomassie Instant Blue and destained by washing with water. The loading of ubiquitin to E2 conjugating enzymes is carried out in an identical manner, except that UBE1 (0.17 μM) is mixed with Ubc13 (2.4 μM) or UbcH7 (2.9 μM) prior to incubation with BAY 11-7082[3].

Cells are transfected with siRNA in 96-well microtiter plates and then cultured for 72 hours in complete NSCLC medium, treated with BAY 11-7082 for 12 hours. Cells are incubated with [3H]thymidine for 3 hours. The cells are collected on filters using an automatic cell harvester and radioactivity on the filters is measured by β-scintillation counting.(Only for Reference)