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To assess whether RT probe binds to any proteins or reacts with free cysteine residues in some proteins that may interfere with the final ratio readouts, we incubated RT with cell lysate, which was dialyzed using a 3 K cut-off membrane to remove all small molecules including GSH, and observed no significant changes in fluorescent signal ratios even at the highest lysate concentration shown (protein concentration is 36 mg ml 1 based on a BCA assay), suggesting that there is no non-specific interactions or reactions between cellular proteins and RT which could significantly change the ratio readouts (Fig

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The characteristics of ZO and JAM proteins are shown in Table 2

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