Ghosh B, Ganea G R, Denson L A, Iannucci R, Jacobs H C, Bogue C W
Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Pediatr Res. 2000 Nov;48(5):634-8. doi: 10.1203/00006450-200011000-00014.
A polyclonal antibody against a glutathione S:-transferase fusion protein containing the 76 COOH-terminal amino acids of Hex, a divergent homeobox gene, was raised in rabbits. Western blot and immunofluorescence reveal that Hex is a 35-37-kD soluble protein present both in the nucleus and cytoplasm of transfected and nontransfected cultured cells as well as in whole mouse embryo. Confocal microscopy of whole mount immunostained mouse embryos at E7. 5 and E8.5 demonstrates that Hex is differentially localized in the cytoplasm and nucleus of definitive endoderm, developing blood islands, and hepatic diverticulum. In particular, in the region of the foregut that gives rise to the liver, Hex expression is nuclear in the endodermal cells of the hepatic diverticulum, whereas expression is primarily cytoplasmic in cells lateral to the liver-forming region. This suggests that nuclear localization of Hex is involved in early hepatic specification and that compartmentalization of Hex protein plays an important role in its function during mouse development.
针对一种谷胱甘肽S-转移酶融合蛋白制备了多克隆抗体，该融合蛋白包含一种差异同源异型框基因Hex的76个羧基末端氨基酸，制备过程是在兔子体内进行的。蛋白质免疫印迹法和免疫荧光法显示，Hex是一种35 - 37千道尔顿的可溶性蛋白，存在于转染和未转染的培养细胞的细胞核和细胞质中，也存在于整个小鼠胚胎中。对E7.5和E8.5的全胚免疫染色小鼠胚胎进行共聚焦显微镜检查表明，Hex在确定内胚层、正在发育的血岛和肝憩室的细胞质和细胞核中存在差异定位。特别是，在形成肝脏的前肠区域，Hex在肝憩室的内胚层细胞中表达于细胞核，而在肝脏形成区域外侧的细胞中，表达主要位于细胞质。这表明Hex的核定位参与早期肝脏特化，并且Hex蛋白的区室化在小鼠发育过程中其功能发挥中起重要作用。
