Mitoq

Zalcitabine (S1719), erastin (S7242), sulfasalazine (S1576), IKE (S8877), MitoQ (S8978), bafilomycin A1 (S1413), staurosporine (S1421), and CCCP (S6494) were purchased from Selleck Chemicals. MitoTEMPO (SML0737) was purchased from Sigma-Aldrich. The antibodies to STING1 (13647), TOMM20 (42406), MFN1 (14739), OPA1 (67589), PINK1 (6946), PRKN (4211), ACTB (3700), CALR (12238), and MAP1LC3B (3868) were purchased from Cell Signaling Technology. The antibody to MFN2 (ab205236) was purchased from Abcam. The antibody to mitHsp70 (MA3-028) was purchased from Thermo Fisher Scientific.

CMT-U27 and CF41.Mg, epithelial-like rounded cells derived from canine mammary gland carcinoma, were procured from ATCC (Manassas, VA, USA). These cells were cultured in Roswell Park Memorial Institute 1640 medium for CMT-U27 cells or Dulbecco’s Modified Eagle’s Medium for CF41.Mg cells, supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin, in a humidified atmosphere containing 5% CO₂ at 37 °C. MitoQ was obtained from Selleck Chemicals LLC (Houston, TX, USA) and diluted in dimethyl sulfoxide (Sigma Aldrich, Saint Louis, MO, USA) to prepare a 1 M stock solution. This stock solution was further diluted to the desired concentration using cell culture medium prior to cell treatment.