Skip to main content
. 2021 Sep 7;46:102126. doi: 10.1016/j.redox.2021.102126

Fig. 1.

Fig. 1

Fluorescent detection of nitro-alkylated proteins in living cells. (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the alk-9-NO2-OA probe 8 and azide-rhodamine. CuAAC, copper-catalyzed azide-alkyne cycloaddition. (B) Time-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells at 2.5 μM alk-9-NO2-OA. (C) Dose-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells after treatment with indicated concentrations of alk-9-NO2-OA for 1 h. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293T cells at 2.5 μM alk-9-NO2-OA for 1 h. 3M and 7M refer to mutants with 3 and 7 cysteines mutated, respectively. (E) Fluorescent detection of nitro-alkylated proteins in different human cell lines treated with 10 μM alk-9-NO2-OA for 15 min. Anti-HA, anti-GAPDH, anti-tubulin blots and Coomassie stains act as loading controls for the accompanying fluorescence gels. Selected protein molecular weight markers are indicated for each gel.