Modulation of 4T1 Breast Cancer Cells

Murine BC 4T1 cell line was maintained and cultured as our previous reports (7 (link), 13 (link)). Sax (0, 0.2, and 0.4 μM) and Sit (0, 0.6, and 1.2 μM) (MCE, Houston, USA), NRF2 specific inhibitor ML-385 (0, 5, and 10 µM) (MCE, Houston, USA), Heme oxygenase 1 (HO-1) specific inhibitor HO-1-IN-1 hydrochloride (0, 5, and 10 µM) (MCE, Houston, USA), reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) (0, 2.5, and 5 mM) (AbMole, Houston, USA), and NRF2 activator alpha-lipoic acid (ALA) (0, 40, and 60 μM) (Dandong Yichuang Co, China) were used as described previously (7 (link)). pNF-кB-TA-luc reporter plasmids were obtained from Beyotime, Jiangsu, China, as described previously (14 (link)). Granulocyte-macrophage colony-stimulating factor (GM-CSF) was purchased from PeproTech, Cranbury, USA. NLRP3 inhibitor MCC950 (0, 5, and 10 µM) (MCE, Houston, USA), caspase-1 inhibitor VX-765 (0, 10, and 20 µM) (AbMole, Houston, USA), and nuclear factor kappa B (NF-κB) specific inhibitor BAY 11-7082 (0, 2, and 4 µM) (Selleck, Houston, USA) were used in this study. For pharmacological intervention assays, 4T1 cells were pretreated with Sax (0.4 μM) or Sit (1.2 μM) for 16 h and then cotreated with indicated inhibitors or activators for additional 4–6 h unless otherwise specified. All chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise indicated.