Confirmation of successful inactivation of the CD59a gene.

(A) Northern blot analysis of CD59a (panel Ai) and CD59b (panel Aii) mRNAs from wild-type (lanes 1,3,5) and knockout (lanes 2,4,6) mouse heart (lanes 1-2), kidney (lanes 3-4), and testis (lanes 5-6). Equal RNA loading is indicated in panel Aiii. The membrane was first hybridized with a 370-bp CD59a cDNA fragment17 (panel Ai), and then stripped and rehybridized with a CD59b-specific probe (panel Aii). Positions of the 18S and 28S ribosomal RNA bands in Ai and Aii are indicated by two horizontal lines. (B) RT-PCR analysis of RNAs from wild-type (WT) and knockout (KO) mouse testis and heart. Primer combinations for CD59a mRNA were as indicated, and 2 primers, cd59b-1 and cd59b-2, were used for CD59b mRNA (primer abbreviations and sequences appear in “Materials and methods”). (C) FACS analysis of CD59a expression on wild-type, heterozygous, and knockout mouse erythrocytes. Wild-type cells stained only with secondary antibody are represented by shaded area.