Fig 2: Analyses of survival-related FRGs in OC. (A) Ferroptosis pivot genes, namely, SLC7A5, ACSL4, XCT, GPX4, and ß-actin protein expression in platinum-resistant tissues (R1-R3) and platinum-sensitive tissues (S1-S3). (B) SLC7A5, ACSL4, XCT, GPX4, and ß-actin protein expression in ovarian cell lines (OVCAR3 and SKOV3) and platinum-resistant OC cell lines (OVCAR3-CDDP and SKOV3-CDDP). (C) Cell viability assays under cisplatin treatment; SKOV3/CDDP showed higher cisplatin tolerance, representing the resistance phenotype. Cells were treated with cisplatin (2.2 µg/ml for SKOV3 and 6.9 µg/ml for SKOV3-CDDP) in the absence or presence of erastin (10 µM), RSL3 (0.1 µM), and NAC (1 mM) for 24 h and cell viability. (D) Glutathione (GSH) (E) and lipid peroxidation (MDA) assays (F) were performed. (G) XCT, GPX4, and ß-actin protein expression in SKOV3 (i) and SKOV3-CDDP (ii) under the abovementioned treatment. Error bars represent the standard deviation (s.d.) of triplicate measurements. *p < 0.05 and **p < 0.01.

Fig 3: The expression levels of the NLRP1 in HTR-8/SVneo cells transfected with siRNA-NLRP1 were detected by western blot (A) and qRT-PCR (B). The expression levels of NLRP1 at the mRNA levels were significantly down-regulated after siRNA-NLRP1 (**P < 0.01), as compared with the H2O2 and siRNA-NC groups. (C–E) Ferroptosis-related protein expression in HTR-8/SVneo cells were detected by western blot. (C) GPX4, (D) TFR1, and (E) ACSL4 (**P < 0.01). (F–H) mRNA expression in HTR-8/SVneo cells was detected by qRT-PCR. (F) GPX4, (G) TFR1, and (H) ACSL4 (*P < 0.05 and **P < 0.01). (I–K) Ferroptosis levels in the HTR-8/SVneo cell model of oxidative stress l. (I) GSH, (J) MDA, (K) SOD, and (L) GPx. (M) CCK-8 assay was used to detect cell activity (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001). Values represent means ± SD, n = 3 per group.

Fig 4: ACSL4 partially localises to MAM. MAM, cytoplasmic and mitochondrial fractions, were prepared from MCF-7 cells. Equal volumes of samples from each MCF-7 fraction were subjected to SDS-PAGE separation and immunoblotted for ACSL4, calnexin, caveolin, HMGCR and Sigma1R. Each panel represents the results of an experiment repeated twice with similar results

Fig 5: Overexpression of BECN1 in HK-2 cells can exacerbate the ferroptosis of cells induced by oxalate. The concentration of oxalate was 2 mmol/L, and the intervention time was 24 hours. Among the four groups of cells, we compared the levels of BECN1 protein relative to GADPH levels (a), the MDA content (b), the GSH content (c), the Fe2+ content (d), the ROS levels (e), the mitochondrial membrane potential (g), GPX4 protein level of the ctrl Ox and Ox+BECN1 groups (h), and the expression of BECN1 and GPX4 mRNA in the four groups of cells relative to that of GAPDH (f). We compared the levels of P62, LC3 II, ACSL4, TFR1, TFH1, GPX4, and xCT proteins in the four groups of cells relative to the GAPDH levels (i). The fluorescence images were all taken under a 200× field of view under an automatic microscope. Data are presented as the means ± SEM from three independent experiments. #P < 0.05, ##P < 0.01, ###P < 0.001, and ####P < 0.0001 versus the ctrl Ox group; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 versus the ctrl NC group. ns: not significant.