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. 2015 Apr 29;56(4):2553–2567. doi: 10.1167/iovs.14-16298

Figure 2.

Figure 2

The β-Gal fragment complementation assay for a HTS of P23H mutant opsin translocation (top left). (A) Immunostaining of U2OS (PLC-EA and P23H-opsin-PK) stable cells treated with 0.1% DMSO (left) and 5 μM 9-cis-retinal (right). PLC-EA is the PH domain of phospholipase C δ fused with the large subunit of β-Gal; and P23H-opsin-PK is the P23H mutant opsin fused with the small subunit of β-Gal. Yellow, P23H-opsin-PK; blue, DAPI. (B) Dose-response curve of 9-cis-retinal assessed by the β-Gal fragment complementation assay. Concentrations of 9-cis-retinal tested were 0.001, 0.005, 0.01, 0.05, 0.10, 0.25, 0.5, 0.75, 1.0, 2.5, 5, 7.5, 10, 15, and 20 μM. Error bars, SDs of triplicate determinations. The HTS quality control parameters (S/B ratio and Z' described in Methods and the EC50 of 9-cis-retinal are listed in the inset.