iTRAQ labeling

Equal amounts of tryptic peptides from TM-treated cells with different time points (dynamic set), as well as from control and triplicate TM-treated OVCAR-3 cells (validation set) were labeled by 8-plex iTRAQ reagents according to the manufacturer’s protocols for protein quantification (dynamic set: 0 h TM treatment: iTRAQ 113 and 119; 6 h treatment: iTRAQ 114; 12 h treatment: iTRAQ 115; 24 h treatment: iTRAQ 116; 48 h treatment: iTRAQ 117 and 121; 72 h treatment: iTRAQ 118; validation set: control: iTRAQ 113 and 117; no. 1 treated cells: iTRAQ 114 and 118; no. 2 treated cells: iTRAQ 115 and 119; no. 3 treated cells: iTRAQ 116 and 121). The iTRAQ-labeled samples were pooled and purified by SCX and Cl8 columns for further analysis.