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Biochemical examination of disulfide bond formation between cysteine pairs in the paddle motif. (a and b) Western blots of purified recombinant wild-type Shaker protein and I325C I364C (a) or (b) T329C L361C double-cysteine mutant proteins prepared under reducing or non-reducing conditions and with or without TEV digestion. All tested proteins contain an N-terminal Flag epitope with or without a TEV site in the S3–S4 linker. Molecular weight standards (MWS) run in the leftmost lane.
