Xenbase Image ID: 144727

Figure 7. Effects of rhodopsin sequence, expression system, and 11-cis-retinal on proteolytic cleavage of P23H rhodopsin. A, Western blot probed with mAb 1D4 showing samples of nontransgenic X. laevis eye extract (a) and extracts from eyes expressing wild-type bovine rhodopsin (b), bovine P23H rhodopsin (cyclic reared) (c), bovine P23H rhodopsin (dark reared) (d), human wild-type rhodopsin (e), human P23H rhodopsin (F1, mild phenotype, cyclic reared) (f), human P23H rhodopsin (F1, mild phenotype, dark reared) (g), human P23H rhodopsin (F1, severe phenotype, cyclic reared) (h), human P23H rhodopsin (F1, severe degeneration, dark reared) (i), extracts of HEK293S cells expressing bovine P23H rhodopsin (j), or bovine wild-type rhodopsin (k). Lanes a and f–i contain four times as much sample as lanes b–e, and lanes f–k were scanned at a more sensitive setting. HEK293S cells were cultured in the presence of 50 μM 11-cis-retinal. B, Western blot probed with 1D4 showing extracts of HEK293S cells expressing wild-type or P23H bovine rhodopsins cultured in the presence or absence of 50 μM 11-cis-retinal. Arrowheads indicate heterogeneously glycosylated (H), mature glycosylated (M), and truncated (T) rhodopsins. C, Graphical representation of the effects of 11-cis-retinal on the ratio of truncated to mature glycosylated rhodopsins (n = 3 for each condition). Although the overall levels of P23H rhodopsin increased in the presence of 11-cis-retinal, the relative abundance of the truncated product decreased by approximately twofold. Image published in: Tam BM and Moritz OL (2007) Copyright © 2007. This image is reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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