Figure 6.
Klar localizes to the nuclear envelope in a KASH-dependent manner. (A) Klar distribution in ovaries. Wild-type ovaries were fixed and stained by Hoechst 33258 for DNA (blue), antibody Klar-C for Klar (green), and WGA for the nuclear envelope (red). Klar protein localized to the nuclear envelope in nurse cells. (B) Klar distribution in various klar mutants. In class II alleles, Klar was either mislocalized throughout the cytoplasm of nurse cells and oocytes (klarmCD4) or not detectable (klarmBX12). For the class I allele klar1, Klar distribution was similar to the wild type. Klar and DNA were detected as for (A). (C) Molecular lesions in class II alleles (left) and their consequence for Klar protein expression by Western analysis (right). klarmCD4 contains a nonsense mutation just before the KASH domain; klarmBX13 and klarmBX12 have chromosomal breaks before or within the KASH domain. A double-headed arrow indicates the approximate location of the Klar-C epitope. By Western analysis, Klar-C detects a 75-kDa band in wild-type ovaries, a shorter form in klarmCD4, and no signal above background for klarmBX12 and klarmBX13. Bars, 8 μm (A) and 40 μm (B).