Figure 8

V2R mutants show limited correction upon F508del-CFTR corrector treatment. Representative blots are shown for V2R V206D (A-C) and V2R L292P (D-F). Representative immunoblot for cells expressing V2R V206D treated with decreasing concentrations of ouabain (ouab), 10nM ABT-888, decreasing concentrations of RDR1 (RDR1), and glafenine (glaf) (A); ABT-888 (ABT), ouabain (ouab), KM60, and latonduine (Lat) (B); KM57 (C). Representative immunoblot for cells expressing V2R L292P treated with decreasing concentrations of ouabain (ouab), KM60, KM57, RDR1 (D); ABT-888 (ABT888), latonduine (Lat), and glafenine (E); carbamazepine (F). In all blots, lanes expressing vector controls (vector), and wild type V2R (wt) are shown. A line above the corresponding lanes indicates where V2R V206D or V2R L292P is expressed. Each blot has a vehicle control (DMSO or DMSO 0.1%) and a positive control (DMSO, 29°C). For V2R V206D, two other positive controls are also included (1% DMSO, glycerol). In (A-C), an arrow indicates core glycosylated V2R protein (core), a bracket indicates complex-glycosylated V2R protein (complex, indicating rescue), an arrowhead indicates nonglycosylated protein (nonglycosylated), and a star indicates GFP. In (D-F), a black arrow indicates mutant core glycosylated monomer, a black bracket indicates mutant complex-glycosylated monomer, and a black arrowhead indicates mutant dimer. The same symbols are also shown in gray to indicate the wild type V2R protein, which has a slightly slower mobility on SDS-PAGE. In all blots tubulin is shown as a loading control.