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S 3. Double labeling of BrdU (sheep anti BrdU) and CK7 (mouse anti CK7) in bladders from 1 month and 6 months old animals. BrdU was developed with DAB and CK7 was developed with vector-red. Bladders from 1 month are shown in a, c, e, and g. Bladders from 6 months were shown in b, d, f, and h. CK7 was expressed specifically in urothelial cells (a and b, original magnification 40x). Negative controls are shown in c, d, e, and f (original magnification 10x). In c and d, primary antibodies were omitted and secondary biotinylated sheep anti mouse was conjugated with vector red. Some background between muscle layer and stroma are present but no influence of CK7 staining for urothelia. In e and f, only vector-red was applied and no background signal present. Double labeling of BrdU and CK7 showing BrdU (g and h, original magnification 63X). Arrows indicate double labeling of BrdU and CK7.