Bw720c

For drug-induced parasite death TaNM1 cells were treated with BW720C at 200 ng/mL for 72 h (Sigma, United States), and as a negative control were treated with an equal volume of DMSO. RNAi experiments were performed in six-well plates. 200 pm negative control or siRNA against target gene were transfected into TaNM1 cells using 10 μL lipofectamine 2000 reagent (Invitrogen, United States) and cells were collected in 36 h after siRNA transfection. All collected cells were used to extract total RNA or protein for RT-qPCR or western blot detection.

Theileria annulata-infected cells TaNM1 were cultured in 12-well culture plates at 10⁵ cells per well and treated with 200 ng/ml (stock 2 mg/ml in ethanol) buparvaquone (BW720c, Sigma). Meanwhile, cells were treated with the same amount of ethanol that used for dissolve BW720c as the control group. HeLa cells were also treated with the same concentration of BW720c to test its effect on the protein level of telomerase reverse transcriptase.
5 × 10⁵ TaNM1 cells were treated with 30 nmol/ml BIBR, 300 nmol/ml novobiocin, and 1 nmol/ml Geldanamycin (MedChemExpress), respectively. The equal volume of DMSO was used to treat the cells as control group.