Skip to main content
. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: Kidney Int. 2012 Apr 11;82(3):278–291. doi: 10.1038/ki.2012.102

Figure 3. Generation and characterization of “humanized” HO-1 BAC (hHO-1 BAC) mice.

Figure 3

(A) Five transgenic mice were obtained from a pair of HO-1+/− BAC positive mice. Primers for HO-1 BAC (a and b, Table 1, Figure 1A) were used to screen whether they were positive for HO-1 BAC. Mice were also genotyped using a combination of primers for the detection of mouse HO-1 gene (lower 160-bp band) and/or neomycin cassette (upper 280-bp band) inserted in order to generate HO-1−/− mice (Table 1). (B) Genomic DNA was used to evaluate the presence of mouse and human HO-1. While the genotyping of HO-1+/+ mice reveals a single band at 160 bp for mouse HO-1, the absence of mouse HO-1 in hHO-1 BAC mice gives a single band at 280 bp denoting the presence of the neomycin cassette derived from the HO-1−/− mice. A heterozygote would reveal two bands at 160 and 280 bp. The presence of human HO-1 in hHO-1 BAC mice is confirmed by using a human specific primer that yields a 320 bp band. (C) Microsomal fractions of kidney and spleen from HO-1+/+, HO-1−/−, and hHO-1 BAC mice were isolated and analyzed by immunoblot for HO-1 expression. The blot was stripped and reprobed with anti-actin antibody for a loading control.