Each caveola contains multiple glycosyl-phosphatidylinositol-anchored membrane proteins.
Each caveola contains multiple glycosyl-phosphatidylinositol-anchored membrane proteins.
复制标题
每个小凹包含多个糖基磷脂酰肌醇锚定膜蛋白。
DOI：
10.1101/sqb.1992.057.01.065
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Rothberg,KG
中科院分区：
文献类型：
--
作者：
Ying,YS;Anderson,RG;Rothberg,KG
方法细胞培养。绒毛膜癌细胞株(ATCC-HTB 36)和神经母细胞瘤细胞株(神经母细胞瘤细胞株)(ATCC-CCL 131)在添加10%胎牛血清和100单位/毫升青霉素/链霉素的改良Eagle‘s培养液中连续单层生长。小鼠成纤维细胞系NIH-3T3(ATCCCRL 1659)和3T3-L1(ATCC-CL 173)细胞在含有4500 mg/1d-葡萄糖、10%小牛血清和100U/ml青霉素/链霉素的改良Eagle‘s培养液中生长。每个实验的细胞都是根据标准方案设置的。第0天，将0.5×104~2.5×104细胞接种于35 mm培养皿中，培养3~5d。2天后更换培养液。为了诱导3T3-L1细胞向脂肪细胞分化，细胞以较高的密度培养，每皿5×104个细胞，培养5d，使细胞后融合。然后在含有胰岛素(10/g/ml)、地塞米松(0.3/ZM)和异丁基甲基黄嘌呤(0.5~M)的完全培养液中诱导2d。去掉地塞米松和异丁基甲基黄嘌呤，细胞在胰岛素(10/xg/ml)中维持。诱导后第8天取材。
MethodsCell culture. JEG-3 cells, a chorionic carcinoma cell line (ATCC-HTB 36), and Neuro-2A, a neuroblastoma cell line (ATCC-CCL 131), were grown continuously as a monolayer in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 100 units/ml penicillin/streptomycin. NIH-3T3 (ATCCCRL 1659) and 3T3-L1 (ATCC-CL 173) cells, mouse fibroblast cell lines, were grown in Dulbecco's modified Eagle's medium containing 4500 mg/1 d-glucose, 10% calf serum, and 100 units/ml penicillin/streptomycin. Cells for each experiment were set up according to a standard protocol. On day 0, 0.5 x 10 4 to 2.5 X 10 4 cells were plated onto 35-mm dishes and grown for 3-5 days. Medium was changed after 2 days. For induction of 3T3-L1 cell differentiation to adipocytes, cells were plated at a higher density, 5 x 104 cells/dish, and grown for 5 days so that cells were postconfluent. Cells were then induced in complete medium containing insulin (10/Lg/ml), dexamethasone (0.3/ZM), and isobutyl methylxanthine (0.5~ M) for 2 days. The dexamethasone and isobutyl methylxanthine were removed and the cells were maintained in insulin (10/xg/ml). Cells were used 8 days after induction.