Fig. 3: BID is covalently modified by ITC probes on N-terminal cysteines. | Cell Death Discovery

Fig. 3: BID is covalently modified by ITC probes on N-terminal cysteines.

From: Quantitative chemical proteomics reveals that phenethyl isothiocyanate covalently targets BID to promote apoptosis

Fig. 3

a In-gel fluorescence validation of BID labeling by ITC probes. b Investigation of the modification sites in BID labeled by PEITC-yne. c Characterization of cysteine residues in BID that are modified by BITC-yne. d Characterization of cysteine residues in BID that are modified by PEITC-yne. e Characterization of cysteine residues in BID that are modified by SFN-yne. For (a–e), HEK293T cells were transfected to express BID or its mutants, labeled with the probe (20 µM) for 0.5 h, and subjected to immunoprecipitation (IP), followed by conjugation with azido-rhodamine and in-gel fluorescence analysis. f Labeling of recombinant BID by ITC probes in vitro. Purified BID protein was incubated with ITC probes (20 µM) for 2 h, conjugated with azido-rhodamine, and analyzed by in-gel fluorescence. Rho: rhodamine; CBB: Coomassie Brilliant Blue staining.

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