Nunnally M H, D'Angelo J M, Craig S W
J Cell Biol. 1980 Oct;87(1):219-26. doi: 10.1083/jcb.87.1.219.
Affinity-purified rabbit antibody to purified chicken gizzard filamin was used in indirect immunofluorescence to localize filamin in dividing chick embryo cells. The antibody was shown to bind only chick embryo cell filamin when whole cell extracts were analyzed by the sensitive sodium dodecyl sulfate-polyacrylamide gel electrophoresis overlay technique described by Adair et al. (1978, J. Cell Biol. 790:281-285). The results show that filamin is located in stress fibers and membrane ruffles during interphase. As cells prophase, the condensing chromosomes are surrounded by diffuse antifilamin staining. No stress fibers are apparent. During metaphase and anaphase, the staining is bright but diffuse. There is often peripheral membrane staining. Filamin is not concentrated in the spindle region but neither is it excluded from the spindle. During cytokinesis, filamin is found highly concentrated in the cleavage furrow in 16 out of 100 cells examined. This frequency of concentration in the furrow is comparable to that observed for alpha-actinin (14%). Myosin concentration in the furrow is more frequent; it is observed in 37% of the cells examined. Neither myosin, alpha-actinin, nor filamin is observed concentrated in the furrow 100% of the time. We conclude that the results are consistent with, but not sufficient to prove, the hypothesis that alpha-actinin and filamin are essential components of the mechanism of cytokinesis.
用亲和纯化的抗纯化鸡砂囊细丝蛋白兔抗体进行间接免疫荧光，以在分裂的鸡胚细胞中定位细丝蛋白。当通过Adair等人（1978年，《细胞生物学杂志》790:281 - 285）描述的灵敏的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳覆盖技术分析全细胞提取物时，该抗体仅与鸡胚细胞细丝蛋白结合。结果表明，细丝蛋白在间期位于应力纤维和膜皱褶中。细胞进入前期时，浓缩的染色体被弥漫性的抗细丝蛋白染色所包围。此时没有明显的应力纤维。在中期和后期，染色明亮但呈弥漫性。常常有周边膜染色。细丝蛋白不集中在纺锤体区域，但也不排除在纺锤体之外。在胞质分裂期间，在所检查的100个细胞中有16个细胞中发现细丝蛋白高度集中在分裂沟中。这种在沟中集中的频率与观察到的α - 辅肌动蛋白的频率相当（14%）。肌球蛋白在沟中的集中更为常见；在所检查的37%的细胞中观察到。肌球蛋白、α - 辅肌动蛋白和细丝蛋白都不会100%的时间都集中在沟中。我们得出结论，这些结果与α - 辅肌动蛋白和细丝蛋白是胞质分裂机制的重要组成部分这一假设一致，但不足以证明该假设。
