Anti gpx4

Sudan Black, Prussian Blue, ( ±) α-LA, Luperox® DI (tert-Butyl peroxide), anti-fatty acid synthase (FAS), and trypsin were purchased from Sigma Chemical Co. (St. Louis, MO). BODIPY® 598/591 C11, MitoTracker Deep Red FM, DAPI, were purchased from Invitrogen/Molecular Probes (Eugene, OR). MitoPeDPP® was purchased from Dojindo Molecular Technologies, Inc. (Rockville,MD) Anti-PANK2, anti-MTND1, anti-NDUFA9, anti-NFS1, anti-ISCU, anti-LYRM4anti-NRF2, PDH hand complex I activity kit and aconitase kit were purchased from Abcam (Cambridge, UK), Anti-mitochondrial 10-formyltetrahydrofolate dehydrogenase (ALDH1L2), anti-alpha-aminoadipic semialdehyde synthase (AASS), anti-FOXN4, anti-hnRNPA/B,anti-NF-Y, anti-Tau, anti-GPX4 and anti-AASDHPPT were purchased from Thermo-Fisher (Waltham, MA). Anti-lipoic acid was acquired from Merck (Darmstadt, Germany). Anti-PLA2G6 and anti-SOD were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-actin was acquired from MyBiosource (San Diego, California, USA). OxyBlot Protein Oxidation Detection Kit was acquired from Merck (Darmstadt, Germany). A cocktail of protease inhibitors (complete cocktail) was purchased from Boehringer Mannheim (Indianapolis, IN). The Immun Star HRP substrate kit was from Bio-Rad Laboratories Inc. (Hercules, CA).

Cells were collected, washed, and lysed in radioimmunoprecipitation assay buffer (Beyotime, China) containing a protease inhibitor cocktail. Cell lysates (20 μg/lane) were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes. Membranes were then kept in 5% skim milk for 1 h and incubated overnight at 4℃ with primary antibodies. Following three washes, the membranes were subjected to incubation with a secondary antibody at room temperature for a duration of 2 h. Visualization was performed using the ECL Plus Ultra-sensitive Chemiluminescence Substrate (Solarbio, China) and a fluorescence and chemiluminescence imaging system (Clinx, China). The primary antibodies used were anti-β-actin (Abcam, UK), anti-GPX4 (Thermo Fisher, USA), anti-SLC7A11 (Thermo Fisher), anti-SCD1 (Abcam), and anti-STK11 (Abcam). The secondary antibody used was goat anti-rabbit IgG H&L (HRP) (Abcam).