Smo:46 nM (TM3Hh12 cells)

方法：转基因果蝇，在眼部表达 APP-C99-Gal4 和 UAS-GRIM(γ-分泌酶切割后激活 GRIM，导致眼粗糙)，果蝇从幼虫至成体在含 Cyclopamine 药(100 nM)食物上饲养，成体后 24 小时内观察。 结果：Cyclopamine 处理组果蝇的粗糙眼表型严重程度显著降低，在体内也能减少γ-分泌酶介导的 APP 切割。[2]

方法：从 C57BL/6N 小鼠分离的原代肝细胞，加入 Cyclopamine （10 μM）和雷帕霉素 （50 nM），处理 24 小时，Seahorse XF 分析仪检测线粒体耗氧率（OCR），计算基础呼吸、最大呼吸、ATP 产量等参数。 结果：Cyclopamine 与雷帕霉素联合显著降低最大呼吸和 ATP 产量。[1] 方法：HeLa 细胞中加入 Cyclopamine （5 μM），处理 24 小时，Western blot 检测 cathepsin D 的成熟形式（31 kDa）与未成熟形式（53 kDa）的比值。 结果：Cyclopamine 处理导致成熟/未成熟 cathepsin D 比值显著降低，表明溶酶体成熟受损。[2]

DMSO:4.12 mg/mL (10.01 mM)

Cells were cultured in triplicate in 96-well plates in assay media to which 5E1 monoclonal antibody, ShhNp and/or cyclopamine were added at 0 h at concentrations indicated in the main text. Viable cell mass was determined by optical density measurements at 490 nm (OD490) at 2 and 4 days using the CellTiter96 colorimetric assay. Relative growth was calculated as OD (day 4) 2 OD (day 2)/OD (day 2) [3].

A total of 0.1 ml Hanks' balanced salt solution and matrigel (1:1) containing 2 × 10^6 cells were injected subcutaneously into CD-1 nude mice. Tumours were grown for 4 days to a minimum volume of 125 mm3; treatment was initiated simultaneously for all subjects. Mice were injected subcutaneously with vector alone (triolein:ethanol 4:1 v/v) or a cyclopamine suspension (1.2 mg per mouse in triolein: ethanol 4:1 v/v) daily for 7 days. At the end of the treatment period, tumours were excised from mice, weighed and then fixed for 3 h at 4 °C with 4% paraformaldehyde, embedded in paraffin wax and sectioned (6 μm). Apoptotic cells were identified by TUNEL using recombinant Tdt as previously described29. Sections were then counterstained with eosin. Eight ×20-magnified fields from regions corresponding to the exterior, middle and interior of two control and two cyclopamine-treated tumours were chosen at random [5].