Ab86707

The total cell lysis was obtained by adding radioimmunoprecipitation assay (RIPA) lysis buffer (Bio-Rad Laboratories, Inc.) to cells. For the pancreatic tissues, they were washed and homogenized in RIPA buffer. After measuring the protein concentrations by using a bicinchoninic acid Protein Assay Kit (Bio-Rad Laboratories, Inc.), the proteins were separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by blotting on polyvinylidene difluoride membrane (Millipore Corp., Bedford, MA, USA). Then, the membranes were blocked with 5% BSA, incubated with the primary and secondary antibodies, and developed using an ECL system (Bio-Rad Laboratories, Inc.). Details of the primary antibodies are as follows:
Anti GPX4, Abcam, Ab125066;
Anti NCOA4, Abcam, Ab86707;
Anti β-actin, Abcam, Ab8226.

Radioimmunoprecipitation assay buffer (RIPA) was applied to lyse cells. Total proteins were then harvested and quantified with bicinchoninic acid assays (Beyotime, Shanghai, China). The target proteins were separated through 10% SDS-PAGE and transferred to polyvinylidene fluoride membranes (Merck Millipore, Burlington, MA, USA). The membranes were blocked with nonfat milk, incubated with primary antibodies, and then incubated with secondary antibody diluted at a ratio of 1:10,000 (Jackson ImmunoResearch, West Grove, PA, USA). The primary antibodies were anti-LC3B (2775, Cell Signaling Technology, Beverly, MA, USA), anti-p62 (88588, Cell Signaling Technology), anti-HuR (ab28660, Abcam, Cambridge, MA, USA), anti-SFRS1 (ab133689), anti-FMRP (ab17722), anti-ALKBH5 (ab195377), anti-IGF2BP1 (ab82968), anti-LIN28A (ab46020), anti-FTH1 (ab65080), anti-NCOA4 (ab86707), anti-BCL-2 (ab32124), anti-BECN1 (ab62557), and anti-GAPDH (ab9485). The protein signals were visualized with enhanced chemiluminescence detection reagents (ECL, Millipore, Burlington, MA, USA) and quantified with Image Lab software (Bio-Rad, Hercules, CA, USA).

Dihydroquercetin (DHQ) (Cat#480-18-2, purity above 98 percent) was purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Erastin (Cat#E7881) and rapamycin (Cat#53123-88-9) were purchased from Sigma Aldrich (Billerica, MA, United States). The SiO₂ (Cat#S5631) particles (approximately 80% diameter: 1–5 μM) were purchased from Sigma-Aldrich and were filtered by sedimentation in accordance with the law of Stokes, subjected to acidic hydrolysis, and baked throughout the night (200°C, 16 h). The SiO₂ particles were prepared for use in both in vivo and in vitro experiments. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (#BS350B) as well as trypan blue exclusion assay kit (#BL707A) were obtained from Biosharp (Anhui, China). Antibodies against GPX4 (ab125066), ferritin heavy chain 1 (FTH1) (ab65080), NCOA4 (ab86707), and microtubule-related protein 1A/1B-light chain 3 (LC3) (ab192890) were all purchased from Abcam Technology (Abcam, Cambridge, United Kingdom). Antibodies against fibronectin (#26836), collagen I (#72026), alpha-smooth muscle actin (α-SMA) (#19245), and GAPDH (#5174), were acquired from Cell Signaling Technology Inc. (Danvers, MA, United States). A secondary antibody was obtained from Proteintech Biotechnology (Wuhan, China).

We purchased renal cancer cell lines ACHN from the American Type Culture Collection (Manassas, VA, USA). ACHN cells were cultured in Eagle’s Minimum Essential Medium enriched with 10% Fetal Bovine Serum (FBS) along with 1% penicillin/streptomycin under 37°C along with 5% CO₂ conditions.
G-CLONE supplied the small chemicals sorafenib (GS0220) along with sodium nitroprusside (GN0200).
Sigma-Aldrich provided us with rapamycin (V900930). ABCAM supplied antibodies to ACSL4 (ab227256), beta-tubulin (ab6046), Beclin-1 (ab210498), LC3 (ab192890), FTH (ab183781), ATG12 (ab155589), and NCOA4 (ab86707). Secondary antibodies linked to HRP, as well as chemiluminescent reagents, were bought from Santa Cruz Biotech (Dallas, TX, USA).
Initially, all chemicals were dispersed in dimethylsulfoxide (DMSO) as a stock solution. The figure legends indicated the chemical treatment duration along with the final levels

The antibodies to ACTB (66009-1-Ig), PARP1 (13371-1-AP), DHODH (14877-1-AP), G6PD (25413-1-AP), and solute carrier family 11 member 2 (SLC11A2, also known as DMT1; 20507-AP) were obtained from Proteintech Biotechnology. The antibodies to SLC25A22 (ab137614), GPX4 (ab125066), solute carrier family 7 member 11 (SLC7A11; ab216876), ACSL4 (ab155282), and NCOA4 (ab86707) were purchased from Abcam. The antibodies to SCD (A16429) were obtained from ABclonal.
The antibodies to AMPKα (5831) and phospho-AMPKα (Thr172; 2535) were obtained from Cell Signaling Technology. RSL3 (S8155), ferrostatin-1 (S7243), liproxstatin-1 (S7699), necrostatin-1 (S8037), Z-VAD-FMK (S7023), erastin (S7242), STS (S1421), OA (S4707), and AICAR (S1802) were purchased from Selleck Chemicals. A939572 (T4515) was purchased from TOPSCIENCE. Oxalacetic acid (328-42-7) was purchased from MACKLIN. GSH monoethyl ester (353905) and NAC amide (A0737) were purchased from MilliporeSigma. BODIPY 581/591 C11 (D3861) was obtained from Thermo Fisher Scientific.