Figure 2.

AGR scanning of the first half of AQP6. (A) Schematic representation for usage of the AGR system with AQP6 (Figure was adapted from Ref.¹⁷) under the terms of the Creative Commons CC BY license. (B) Attaching the N-terminus of AQP6 (comprising residues AQP6^1–10) C-terminally to AGR, results in a construct that reaches the PM (white arrows). (C) In a similar manner, when the first transmembrane domain (TMD) of AQP6 (comprising residues AQP6^7–34) is C-terminally attached to AGR, this construct also reaches the PM. (D) Using the entire AQP6^1–34 residues (which include N-terminus and first TMD), instead of the subunits indicated in (A, B), also results in PM expression when attached to AGR. (E) When residues AQP6^30–42 are used, this construct also localized to the PM. (F) Similarly, when peptide residues AQP6^62–101 are attached to AGR, the construct localized to the PM. Finally, (G) when AQP6^121–138 peptide residues are used the construct also reached the PM. Cells were observed microscopically using an upright Olympus BX51WI equipped with 100 × 1 NA objective and images were captured using a Grasshopper3 CMOS camera (FLIR, Richmond, BC, Canada) controlled by Leica LAS X version 3.5 software (available at: https://www.leica-microsystems.com/products/microscope-software/p/leica-las-x-ls/). All figures were created using Adobe Illustrator CS6 (Available at: https://adobe.com/products/illustrator), under an Adobe Inc., Creative Cloud Desktop 2019 shared device license to Case Western Reserve University (CWRU) that operates until 3/31/2022. Scale bars: 8 µm.