Molecular characterization of gene-trap tTAfunction A). Insertions 02A-0001 in the mouse Car12 gene and insertion 02A-0002 in the mouse ENSMUSG00000066992 gene. Genes are shown in the orientation of transcription from left to right, vertical lines are exons, along with the position and orientation of the T2/GT2/tTA insertion (arrows). B) RT-PCR detection of Car12 (top) and Car12-IRES-tTA chimeric (bottom) transcription in wild type and 02A-0001 carrier tissues. Kd-kidney, Br-brain, Lv-liver, Te-testes, Ad-adipose, Co-colon,SI-small intestine, Ov-ovary, Mu-muscle, Ht-heart C) RT-PCR analysis of gene disruption in tissues from wild type (+/+), heterozygous (+/ins), and homozygous (ins/ins) carriers of insertions 02A-0001 and 02A-0002. C) Western analysis of CA-XII expression in a wild type, heterozygous and homozygous carrier of insertion 02A-0001. Cross-reaction of the CA-XII antibody with other isoforms serves as a loading control (arrowhead).