Fig. 2
From: A network of chaperones prevents and detects failures in membrane protein lipid bilayer integration
Cx32L90H shows defects in gap-junction formation and rapid degradation. a COS-7 cells were transfected with the indicated constructs and immunostained for FLAG-tagged Cx32 (magenta), PDI (yellow) as an ER marker, or GM130 (yellow) as a Golgi marker. Nuclei were stained with DAPI (blue). Anti-FLAG immunofluorescence data are depicted as maximum intensity projections from deconvoluted z-stacks, while PDI, GM130, and nuclei are shown as a central cell plane from the same, nondeconvoluted images. Gap-junction plates lining cell–cell boundaries are indicated with white arrowheads. Pictures are representative of cells from at least three different biological replicates. Scale bars correspond to 20 µm. b HEK293T cells transfected with the indicated constructs were incubated with either cycloheximide (CHX) alone, or additionally with the proteasome inhibitor MG132 where indicated. Arrowheads indicate monomer (M) and dimer (D) bands quantified to determine Cx32 turnover. Quantifications are shown below the immunoblots (mean ± SEM, N ≥ 3). c Half-lifes without MG132 for Cx32wt and Cx32L90H as derived from b are shown
