Immunofluorescent detection of activation of initiator caspases-8 and -9 during pharmacologically induced apoptosis of cultured HeLa and endothelial cells.
Immunofluorescent detection of activation of initiator caspases-8 and -9 during pharmacologically induced apoptosis of cultured HeLa and endothelial cells.
复制标题
在药理诱导培养的 HeLa 和内皮细胞凋亡过程中，用免疫荧光检测启动子 caspase-8 和 -9 的激活。
DOI：
10.1007/s00418-003-0536-1
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发表时间：
2003
影响因子：
2.3
通讯作者：
Sahni,SanjeevK
中科院分区：
文献类型：
--
作者：
Joshi,SureshG;Sahni,SanjeevK
Caspases, a group of cysteine-activated aspartate-directed proteases, play an integral role in the execution of programmed cell death or apoptosis. In the cellular caspase cascade, the processing of native proenzymes into activated forms of downstream, effector caspases is dependent on the activation of initiator caspases-8 and -9. We describe a staining procedure for immunofluorescence-based analysis of activation of caspase-8 and -9 during pharmacologically induced apoptosis in primary cultures of human umbilical vein-derived endothelial cells and in an established line of HeLa cells. Using cleavage site-directed antibodies, specific intracellular detection for cleaved fragments of caspase-8 and -9 was accomplished during apoptosis induced by staurosporine and etoposide. The population of cells displaying morphological signs of apoptosis, evidence for DNA strand breaks by TUNEL analysis, and positive staining for active forms of caspase-8 and caspase-9 increased with the duration of treatment, suggesting activation of initiator caspases in correlation with the onset and progression of apoptosis. The application of immunocytochemical staining procedures for quick and specific in situ detection may effectively aid the identification of participating upstream caspases and elucidation of complex apoptosis signaling mechanisms.
