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FIG. 2

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RT-PCR and Western blot analysis. (A to C) Total RNA was isolated from the eyes (A and B) and kidney (C) of 11-cis-RoDH−/− mice (lanes 1 and 5), 11-cis-RoDH+/− mice (lanes 2 and 6), and 11-cis-RoDH+/+ mice (lanes 3 and 7). Primers specific for 11-cis-RoDH, opsin, and actin were used. The 11-cis-RoDH transcript is absent in 11-cis-RoDH−/− mice. Actin and opsin transcripts are present. No products were amplified when template cDNA was omitted (lanes 4 and 8). Lanes M, Clon16.1.1 size markers or 100-bp DNA ladder. (D and E) For Western blot analysis, 5 μg of 1% Triton X-100–PBS-extractable ocular protein from an 11-cis-RoDH+/+ mouse, 11-cis-RoDH+/− mouse, and 11-cis-RoDH−/− mouse was electrophoresed on an SDS–13.7% polyacrylamide–tricine gel and transferred to nitrocellulose. Blots were incubated with a monoclonal antibody against 11-cis-RoDH (D) or opsin (E). Sizes are shown in kilodaltons.

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