Fig. 2. In-gel fluorescence detection of lactylated proteins with YnLac. (A) Concentration-dependent metabolic labelling of core histones by YnLac. (B) Dose-dependent competition of core histone labelling by YnLac with sodium l-lactate. (C) Concentration-dependent metabolic labelling of cytoplasmic proteins by YnLac. (D) Dose-dependent competition of cytoplasmic protein labelling by YnLac with sodium l-lactate. HEK293T cells were incubated with YnLac at varying concentrations for 8 hours. The core histones and cytoplasmic proteins were separated for click conjugation with az-rho and in-gel fluorescence analysis. For l-lactate competition, HEK293T cells were pre-treated with sodium l-lactate for 4 hours and co-incubated with YnLac (20 mM) and sodium l-lactate at indicated doses (shown as fold of YnLac concentration) for 8 hours. “Rho” represents the rhodamine fluorescence channel. Coomassie brilliant blue (CBB) staining is included as the protein loading control.