Fig. 9: Dose dependent methionine labeling in live cells: cytotoxicity analysis and protein localization insights.

a Dose dependent labeling in live T-47D cells and cytotoxicity analysis showed less than 15% cell death even with 2 mM concentration of CuNiP reagents. Data are represented as mean ± SD (n = 2 biological replicates). b Lysis, and in-gel fluorescence analysis of T-47D cell lysates obtained by labeling proteins in live T-47D cells using CuNiP clearly showed a dose-dependent modification of proteins. Proteomics analysis of modified lysates also demonstrated a dose-dependent labeling of proteins with 1i (100 µM = 20 proteins; 250 µM = 42 proteins; 500 µM = 229 proteins; 1 mM = 305 proteins; 2 mM = 236 proteins). This experiment was repeated (n = 3 biological replicates) with similar results. Excel sheet of analysis is included as Supplementary Data 4. c Gene ontology analysis of cellular compartment of modified proteins showed a significant modification of membrane-bound proteins, in addition to cytoplasmic proteins within the cell. d Gene ontology analysis of biological processes of modified proteins showed a significant modification of proteins of diverse biological functions. e Confocal imaging of CuNiP-modified T-47D cells (>50 cells) showed the dose-dependent modification of proteins inside cells with a wide spatio-temporal distribution (membrane, cytoplasm, and nuclear proteins). This experiment was repeated (n = 2 biological replicates) with similar results. Source data are provided as a Source Data file. Figure 9, created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license” (Agreement number: YB26NY9L7L).