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

Fig. 4.

Fig. 4.

Isomerohydrolase activity in COS-1 cells expressing both RPE65 and LRAT. (A) Western blot analysis of COS-1 cells coexpressing LRAT and RPE65. Lanes 1, positive control 10 μg of microsomal proteins from bovine RPE (for the RPE65 blot) and 1.25 μg of cell lysates of Sf9 cells expressing LRAT (for the LRAT blot); 2, negative control (50 μg of total proteins from untreated cells); 3, 50 μg of cell lysate of COS-1 cells transfected with the LRAT expression plasmid and infected with Ad-RPE65 at MOI of 40; 4, 50μg of cell lysate transfected with the same amount of the LRAT plasmid and infected with Ad-RPE65 at MOI of 300. (B–F) Retinoids generated after incubation of 250 μg of COS-1 cell lysates with 0.2 μM all-trans [3H]retinol for 1.5 h: untreated cells (B), cells infected by Ad-RPE65 alone (C), cells transfected by the LRAT plasmid alone (D), cells transfected with the LRAT plasmid and infected by Ad-RPE65 at MOI of 40 (E), and cells transfected with the LRAT plasmid and infected by Ad-RPE65 at MOI 300 (F). Peaks 1, retinyl esters; 2, all-trans retinal; 3, 11-cis retinol; 4, 13-cis retinol; 5, all-trans retinol.