Figure 1.

Experimental schemes of the (a) BA and (b) MC experiments. In the BA experiment, cells were treated with 15 μM atorvastatin for 24 hours before harvest. After cell lysis, proteins were reduced, alkylated, and digested. Enrichment was performed at the peptide level with boronic acid-conjugated magnetic beads. Enriched glycopeptides were deglycosylated, fractionated, and subjected to LC-MS/MS analysis. In the MC experiment, 100 μM Ac₄GalNAz was added into both heavy and light cells, and heavy cells were treated with atorvastatin. After cell lysis, the lysate was incubated with 100 μMDBCO-sulfo-biotin, followed by protein reduction, alkylation, and digestion. Enrichment was performed at the peptide level with NeutrAvidin agarose beads.