Osborn M, Weber K
Cell. 1977 Nov;12(3):561-71. doi: 10.1016/0092-8674(77)90257-4.
Monospecific tubulin antibodies have been used in indirect immunofluorescence microscopy on a variety of well characterized, transformed cell lines grown in tissue culture. Networks of colcemid-sensitive fibers are seen in SV40-transformed 3T3 cells, SV40-transformed rat embryo cells, HeLa cells and other transformed cell lines. In each case, greater than 90% of the cells contain visible microtubular networks, and where individual microtubules can be distinguished, they run for long distances. Documentation of these metworks is more difficult in transformed than in normal cells, because transformed cells are in general more rounded and have less well spread cytoplasm. In addition, the microtubular networks can be readily visualized in "cytoskeletons" of both normal and transformed cells, obtained by treatment of cells with nonionic detergents in a buffer which stabilizes microtubules in vitro. Addition of calcium to this buffer results in in situ fragmentation and destruction of the microtubular network. In view of these results, we conclude that transformed cells contain significant numbers of microtubules, and that in transformed cells, as in normal cells, microtubules are arranged in networks.
单特异性微管蛋白抗体已用于对多种在组织培养中生长、特性明确的转化细胞系进行间接免疫荧光显微镜检查。在SV40转化的3T3细胞、SV40转化的大鼠胚胎细胞、HeLa细胞及其他转化细胞系中可见秋水仙酰胺敏感纤维网络。在每种情况下，超过90%的细胞含有可见的微管网络，在能分辨出单个微管的地方，它们延伸很长距离。与正常细胞相比，在转化细胞中记录这些网络更困难，因为转化细胞总体上更圆，细胞质铺展性较差。此外，通过在体外稳定微管的缓冲液中用非离子去污剂处理细胞，可在正常和转化细胞的“细胞骨架”中很容易地观察到微管网络。向该缓冲液中添加钙会导致微管网络原位断裂和破坏。鉴于这些结果，我们得出结论：转化细胞含有大量微管，并且在转化细胞中，与正常细胞一样，微管排列成网络。
