Figure 5
hClC-1 oligomerization is not impaired by Q43R.
(a) SDS-PAGE gel (4–10% gradient) of WT and Q43R hClC-1 after expression in Xenopus oocytes and purification via StrepTactin chromatography. Proteins were visualized by scanning of plasma membrane-bound fluorescence (upper panel) or 35S-labeled total protein (lower panel). (b) Oligomeric state of WT and mutant hClC-1 channels. Indicated hClC-1 proteins were purified via metal affinity chromatography, resolved by BN-PAGE (4–12% gradient) and visualized by scanning of YFP fluorescence (upper panel) and 35S-labeled total protein (lower panel). The protein migration is shown both under native conditions and following partial denaturation after a 1-h incubation with 0.1% SDS at 37 °C, as indicated. The gray ovals without and with yellow balls schematically illustrate YFP-less and YFP-fused hClC-1, respectively, to indicate the native dimeric and denatured protomeric states of the corresponding protein bands. The data shown represent samples run on the same SDS-PAGE gel or BN-PAGE gel; irrelevant lanes were cropped from the figure as indicated by vertical lines. The respective source images of the individual gels are shown in Supplemental Fig. S3a and b.
