Cytoselect 24 well cell migration and invasion assay combo kit

Transwell assay was performed to evaluate cell migration and invasion using a CytoSelect™ 24‐Well Cell Migration and Invasion Assay Combo Kit (CBA‐100; Cell Biolabs) following the manufacturer's instructions. Briefly, cells were resuspended and placed in the upper chamber, while fresh medium containing 30% FBS was added to the lower chamber. Twenty‐four hours later, cells remaining on the upper surface were removed, and then the inserts were washed and incubated with 200 μL extraction solution for 10 min, which was then measured at 560 nm by a Synergy HT microplate reader. For the analysis of cell invasion, the inserts were coated with a uniform layer of dried basement membrane matrix solution.

Ferrostain‐1 (Fer‐1, #S7243, an inhibitor of ferroptosis), liproxstatin‐1 (Lip‐1, #S7699, an inhibitor of ferroptosis), necrostatin‐1 (Nec‐1, #S8087, an inhibitor of necroptosis), Z‐VAD‐FMK (Z‐VAD, #S7023, an inhibitor of apoptosis), chloroquine (CQ, #S6999; an inhibitor of autophagy) and 2′, 7’‐Dichlorodihydrofluorescein diacetate (DCFH‐DA, #S9687) were purchased from Selleck (Houston, TX, USA). Cell Counting Kit 8 (CCK8) (#ab228554), Lactate Dehydrogenase (LDH) Assay Kit (#ab102526), Lipid Peroxidation Assay Kit (#ab118970), 12(S)‐hydroxyeicosatetraenoic acid (HETE) ELISA Kit (#ab133034), 15(S)‐HETE ELISA Kit (#ab133035), Iron Assay Kit (#ab83366) and Glutathione (GSH) Assay Kit (#ab239727) were purchased from Abcam (Cambridge, UK). CytoSelect™ 24‐Well Cell Migration and Invasion Assay Combo Kit (#CBA‐100) was purchased from Cell Biolabs (San Diego, CA, USA).