质量

溶剂体积

浓度

1mM

4.6682 mL

2.3341 mL

体内研究（动物模型）：

动物模型（Animal Model）：Tumor-bearing SCID mice (both s.c. and orthotopically implanted with human pancreatic adenocarcinoma cells PK1)

药物配制（Preparation）：Wortmannin was dissolved at 0.4 mg/ml in DMSO, and diluted with 0.9% NaCl before use.

实验方法（Assay）：A) In the first set of experiments, groups of tumor-bearing SCID mice (n = 4 each) were given single bolus injections of 0.7 mg/kg wortmannin via the tail vein and killed at various time points. B)  In a second series of experiments, groups of animals (n = 4 each) were injected i.v. with increasing doses of wortmannin (0.175, 0.35, and 0.7 mg/kg) and killed at 4 h. C) In a third series of experiments, tumor-bearing SCID mice were randomly assigned to four groups (n = 4 each). Each group was given i.v. bolus injections of either the drug vehicle (≤1% DMSO), 80 mg/kg of gemcitabine for 48 h, 0.7 mg/kg of wortmannin for 4 h, or 80 mg/kg of gemcitabine for 48 h and then 0.7 mg/kg wortmannin for 4 h.

体外研究（细胞实验）：

动物模型（Animal Model）：Hepatocytes from Wistar rats

药物配制（Preparation）：Wortmannin was dissolved in DMSO at 2mM, and stored at -20℃ in the dark. Dilutions in 0.9% NaCl were prepared just before the start of each experiment. The final DMSO concentration in the incubation medium did not exceed 0.25 % (by vol.). This concentration of DMSO did not affect the processes that were tested.

实验方法（Assay）：Electron micrographs of isolated rat hepatocytes that were incubated for 60 min (5 mg dry cells/ml) with 50μM vinblastin alone (A, B) or in the presence of 50μM vinblastin plus 100 nM wortmannin.

结果（Results）：Autophagosomes could hardly be detected in cells that were incubated in the presence of 100 nM wortmannin (Fig. 3 C) compared to cells incubated under maximal autophagic conditions (Fig. 3 A and B).