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. 2014 Sep 3;55(9):5572–5581. doi: 10.1167/iovs.13-13717

Figure 2.

Figure 2

Confirmation of genotype and protein expression of Cyp4v3−/− mice compared to wild-type controls. (a) PCR of wild-type (lanes 2, 7, and 12), heterozygote (lanes 3, 8, and 13), and Cyp4v3−/− mice (lanes 4, 9, and 14) shows that wild-type mice have positive bands for Cyp4v3 (lane 2), negative bands for the LacZ reporter construct (lane 7), and positive bands for FABP to verify the presence of genomic DNA (lane 12). Heterozygotes show bands for all three genes, and Cyp4v3−/− mice are negative for Cyp4v3 (lane 4), positive for the LacZ reporter construct (lane 9), and positive for FABP (lane 14). This confirms that Cyp4v3−/− mice lack functional Cyp4v3, and the targeting vector was successfully incorporated into the genome. (b) Western blot analysis of Cyp4v3−/− mice (lanes 2, 3, and 4), wild-type mice (lanes 5, 6, and 7), and 0.18 pmol human purified CYP4V2 as a validation control. Lower bands are β-actin, used to verify the presence of equivalent total protein in each lane, and upper bands show the presence (wild-type) and absence (Cyp4v3−/−) of CYP4V3 protein expression, as expected. This confirms that mice without functional Cyp4v3 do not produce the corresponding protein.