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. 2005 Aug 22;102(35):12413–12418. doi: 10.1073/pnas.0503460102

Fig. 2.

Fig. 2.

Isomerohydrolase activity in QBI-293A cells expressing RPE65 and LRAT. (A) Western blot analysis of total cell lysates. Lane 1, 10 μg of microsomal proteins from bovine RPE (for the RPE65 blot) and 1.25 μg of cell lysates of Sf9 cell expressing LRAT (for the LRAT blot) as positive controls; lanes 2–5, 50 μg of total proteins from 293A cells infected by Ad-GFP (lane 2), transfected with the LRAT plasmid alone (lane 3), infected with Ad-RPE65 alone (lane 4), and transfected with the LRAT plasmid and infected with Ad-RPE65 (lane 5). (B–E) Retinoids generated after incubation of 250 μg of total cellular protein with 0.2 μM all-trans [3H]retinol for 1.5 h. (B) Cells infected by Ad-GFP. (C) Cells infected with Ad-RPE65 alone. (D) Cells transfected with the LRAT plasmid alone. (E) Cells infected by Ad-RPE65 and transfected with LRAT. (F) 11-cis [3H]retinol standard. Peaks 1, retinyl esters; 2, all-trans retinal; 3, 11-cis retinol; 4, 13-cis retinol; 5, all-trans retinol.