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. 2017 Nov 14;27(2):295–306. doi: 10.1093/hmg/ddx400

Figure 6.

Figure 6.

Purification and functional analysis of p.Asn965Ser (N965S) ABCA4. (A) WT and p.Asn965Ser (N965S) ABCA4 were purified from detergent-solubilized retina membranes by immunoaffinity chromatography on a Rim 3F4 matrix and analyzed on SDS gels stained with Coomassie blue (CB) and western blot labeled with the anti-ABCA4 antibody (Rim 3F4). Approximately, 0.2 µg of WT and 0.1 µg of p.Asn965Ser ABCA4 was applied to the lanes for CB staining and 0.08 µg was applied to the lanes for Western blotting. (B, C) ATPase activity of WT (black) and p.Asn965Ser ABCA4 (grey) purified and reconstituted into phosphatidylethanolamine-containing liposomes in the presence and absence of all-trans retinal. (B). ATPase activity of ABCA4 proteins purified from mouse retinal membranes. Mean ± SD for n = 3. (C) ATPase activity of ABCA4 proteins expressed and purified from HEK293 cells. Mean ± SD for n = 4. ATPase assays were carried out using the same concentration of WT and p.Asn965Ser ABCA4. Data are plotted as a percentage of WT ABCA4 ATPase activity in the absence of retinal.