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. 2019 Nov 13;28(2):367–381. doi: 10.1016/j.ymthe.2019.11.005

Figure 5.

Figure 5

GPR108 N- and C-Terminal Domains Are Required for AAV Entry

(A) Predicted membrane topology of nonfunctional GPR107, with indicated disulfide bond location, furin cleavage site, alanine-glycine linker, and functional GPR108. (B) Rescue of Anc80 transduction by transient transfection with FLAG-tagged GPR107/GPR108 chimeric proteins or transfected empty vector (EV). Cells transduced with 10,000 VG/cell, CMV.Luciferase.SVPA transgene after hAd5 helper virus co-infection. (C) Corresponding anti-FLAG western blot from cellular membrane fractions demonstrating protein expression in cells transduced in (B). Black arrows point to expected full-length protein; gray arrows point to expected glycosylation states; red arrows point to protein aggregates; and yellow arrows point to expected N-terminal domain cleavage product. AIF (apoptosis-inducing factor 1) was used as a membrane protein loading control. (D) Indirect immunofluorescence of chimeric proteins using anti-FLAG antibody as described in Figure 4E.