Identification of a novel mammalian KASH protein. (A) Total testis extracts were loaded, and Western blotting was performed with KASH5 serum. A single band was detected corresponding to the predicted size (72 kD). (B) Chromosome spreads from spermatocytes were stained with KASH5 and CENP-C antibodies. (C) Chromosome spreads from spermatocytes were stained with KASH5 and TRF2 antibodies. Magnified images of the boxed area are shown on the right. (D) Amino acid sequence alignment of KASH proteins. Identical amino acids are shaded in black, and similar amino acids are shaded in gray. Dm, Drosophila melanogaster; Mm, Mus musculus; Ce, C. elegans; Sp, S. pombe. (E and F) mRNA expression in various tissues was analyzed by RT-PCR. E, embryonic day; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. Bars, 3 µm.