**2.4 Cell lysis and protein preparation**

Ivermectin treatment and 0.1% DMSO treatment TOV-21G cells were collected and lysed by protein isolation buffer, respectively. (i) TOV-21G cells collection: A total of 500 μL trypsin was added to each 10-cm culture flasks for several minutes and collected with centrifugation (800 × g, 5 min). TOV-21G cells were washed with ice-cold phosphate buffer solution (PBS) for three times. (ii) Protein isolation buffer: 2 mM thiourea, 4% CHAPS (3-[(3-cholamidopropyl)-dimethylammonio] -1-propane), 7 M urea, 100 mM dithiothreitol (DTT), and 2% ampholyte. (iii) TOV-21G cells lysis: A total of 200 μL protein isolation buffer was added to each 10-cm culture flasks for 30 min (ice-cold) and then oscillated with five vortex cycles. (iv) SILAC-labeled protein collection: Protein isolation buffer was centrifuged (13,000 × g, 20 min, 4°C), and the SILAC-labeled protein samples were collected from the supernatants in new tubes. (v) Protein concentrations measurement: Protein concentrations of the SILAC-labeled protein samples were measured with the 2-D quant protein assay kit (Bio-Rad, US).
