Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Zearalenone-Induced Ferroptosis on Mice Spermatogenesis

doi: 10.3390/ani12213026

Figure Lengend Snippet: List of primers employed for q-RT-PCR.

Article Snippet: The proteins were then transferred onto PVDF membranes (GE Bioscience, Newark, NJ, USA) and then blocked with 5% BSA (dissolved in TBST) for 1 h and the membrane was incubated with anti-SYCP3 (ab97672; Abcam, Cambridge, UK), anti-DDX4 (bs-22896R; Bioss Biotech, Beijing, China), anti-SOX9 (ab185966; Abcam, Cambridge, UK), anti-SLC7A11 (bs-6883R; Bioss Biotech, Beijing, China), anti-GPX4 (bs-3884R; Bioss Biotech, Beijing, China), anti-4-HNE (ab48506; Abcam), and anti-β-actin (bs-0061R; Bioss Biotech, Beijing, China) overnight at 4 °C.

Techniques: Sequencing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Zearalenone-Induced Ferroptosis on Mice Spermatogenesis

doi: 10.3390/ani12213026

Figure Lengend Snippet: Expressions of genes, proteins involved in ferroptosis in testis of mice in each group. ( A ) Nrf2 mRNA expression. ( B ) SLC7A11 mRNA expression. ( C ) GPX4 mRNA expression. ( D ) Western blot detection of SLC7A11, GPX4, and 4-HNE protein. ( E ) Image J analysis showing the grey value of SLC7A11. ( F ) Image J analysis showing the grey value of GPX4. ( G ) Image J analysis showing the grey value of 4-HNE. The data are expressed as means ± SEM, with n ≥ 3; Different lowercase letters (a, b, c, d) indicate significant differences ( p < 0.05).

Article Snippet: The proteins were then transferred onto PVDF membranes (GE Bioscience, Newark, NJ, USA) and then blocked with 5% BSA (dissolved in TBST) for 1 h and the membrane was incubated with anti-SYCP3 (ab97672; Abcam, Cambridge, UK), anti-DDX4 (bs-22896R; Bioss Biotech, Beijing, China), anti-SOX9 (ab185966; Abcam, Cambridge, UK), anti-SLC7A11 (bs-6883R; Bioss Biotech, Beijing, China), anti-GPX4 (bs-3884R; Bioss Biotech, Beijing, China), anti-4-HNE (ab48506; Abcam), and anti-β-actin (bs-0061R; Bioss Biotech, Beijing, China) overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: An eCIRP inhibitor attenuates fibrosis and ferroptosis in ischemia and reperfusion induced chronic kidney disease.

doi: 10.1186/s10020-025-01071-2

Figure Lengend Snippet: Fig. 4 C23 reduced myofibroblast formation and ferroptosis in the kidneys in RIR-induced CKD mice. Renal tissues were sectioned and analyzed by immuno histochemistry for α-SMA (A) and quantified using ImageJ (B). Renal tissue sections were stained using Perl’s for iron deposition assessment. Represen tative photomicrographs at 400x magnification are shown (C). The blue intensity was determined using the imageJ software (D). Protein lysates from the different groups underwent Western Blotting. The blots were scanned, shown as cropped gel images and quantified by densitometry (E). The band intensity of GPX4 was normalized to the corresponding band intensity of β-actin. Protein lysates from the different groups were measured for the levels of lipid peroxidation using a MDA kit (F). Data are expressed as means ± SEM (n = 4–6/group) and compared by one-way ANOVA and the SNK method and using Kruskal-Wallis (non-parametric) test; *P < 0.05 vs. Sham or control; #P < 0.05 vs. RIR-Veh

Article Snippet: After overnight incubation at 4 °C with primary antibodies, including TGF-β1 (1:1,000) (Cell Signaling), GPX4 (1:500) (Cell Signaling), and β-actin (1:10,000) (Sigma-Aldrich, Louis, MO, USA), the membranes were then incubated with an HRP-conjugated secondary antibody (1:10,000) (Amersham, Little Chalfont, UK) for 1 h. The bands were detected with ECL.

Techniques: Immunohistochemistry, Staining, Software, Western Blot, Control

Journal: Oxidative medicine and cellular longevity

Article Title: Melatonin Inhibits the Ferroptosis Pathway in Rat Bone Marrow Mesenchymal Stem Cells by Activating the PI3K/AKT/mTOR Signaling Axis to Attenuate Steroid-Induced Osteoporosis.

doi: 10.1155/2022/8223737

Figure Lengend Snippet: Figure 1: DEX activates the ferroptotic pathway of BMSCs. (a) ROS staining was performed to test the correlation between different concentrations of DEX and the level of oxidative stress. (b) Quantitative analysis of the number of ROS-positive cells per field in (a). (c) Annexin V-mCherry/SYTOX Green detection kit was used to detect cell death. (d) Quantitative analysis of the percentage of SYTOX green-positive cells in (c). (e–i) BMSCs were treated with various concentrations of DEX for 24 h, and the expressions of system xc-, ACSL4, GPX4, and FSP1were analyzed by western blot and qRT-PCR. (j) Images of immunofluorescence staining of GPX4 in BMSCs after treatment with DEX (10-3 M) for 72 h. (k) Quantification of the fluorescence intensity of GPX4 immunofluorescence positively stained cells. These studies were performed at least 3 biological replicates. Data represent mean ± SD (n = 3). ∗P < 0:05, ∗∗P < 0:01, ∗∗∗P < 0:005 compared with control group.

Article Snippet: To more fully locate and quantitatively examine antigenic substances in tissues, sections were first stained with primary antibodies against GPX4 (1 : 200, orb340797, Biorbyt), CD90 (1 : 500,RT1615, 0 20 40 60 80 G PX 4 po sit iv e c el l nu m be r ( m m –2 ) ⁎⁎⁎ Co nt ro l M od el (i) 0 20 40 60 FS P1 p os iti ve ce ll nu m be r ( m m –2 ) ⁎⁎⁎ Co nt ro l M od el (j) 0 20 40 60 M T1 p os iti ve ce ll nu m be r ( m m –2 ) ⁎⁎⁎ Co nt ro l M od el (k) Figure 2: DEX activates the ferroptotic pathway in SIOP. (a) Quantification of the protein level of MT by ELISA. (b) Images of micro-CT. (c) BMD(g/cm3). (d) BV/TV (%). (e) Images of immunofluorescence double staining of CD90 and GPX4 in bone tissues. (f) Quantitative analysis of the area of GPX4/CD90-positive stains. (g) IHC staining of system xc-, GPX4, FSP1, and MT1, and the IHC-positive cells were marked with black arrows. (h–k) Quantitative analysis of the number of the IHC-positive cells in (g).

Techniques: Staining, Western Blot, Quantitative RT-PCR, Control

Journal: Frontiers in cell and developmental biology

Article Title: Targeting of the Lipid Metabolism Impairs Resistance to BRAF Kinase Inhibitor in Melanoma.

doi: 10.3389/fcell.2022.927118

Figure Lengend Snippet: FIGURE 7 | Effects of lipid starvation on ferroptosis in melanoma cell lines. (A). Lipid peroxidation is unaffected by lipid starvation in melanoma cells. Induction of lipid peroxidation as detected by MDA measurement after 24 h culture in standard or in lipid-free medium and after treatment for 24 h with PLX4032 at concentrations corresponding to IC80 as calculated after 72 h exposure (0.3 and 0.89 µM for LM16 and LM36, respectively; 23 and 13.9 µM for LM16R and LM36R, respectively). ***p < 0.0001 by one-way ANOVA followed by Bonferroni correction. Experiments were repeated twice. (B). GPX4 expression levels in lysates obtained from LM16, LM16R, LM36, LM36R cells grown for 48 h in the presence or absence of lipids. β–tubulin was used as loading control. (C). Induction of ferroptosis as detected by reduced GSH/GSSG ratio at 24 h culture in standard or in lipid-free medium after treatment with PLX4032 (3 µM), avasimibe (12.5 µM), alone or in combination. Ctr: untreated control; A: avasimibe; PLX: PLX4032; A + PLX: avasimibe + PLX4032. **p < 0.01, ***p < 0.0001 by one-way ANOVA followed by Bonferroni correction.

Article Snippet: Blots were pre-blocked in PBS containing 5% (w/v) dried no fat milk and then incubated overnight at 4°C with antibodies to FASN (HPA006461, Sigma-Aldrich, St. Louis, MO, United States), DHCR24 (2033, Cell Signaling Technology, Danvers, MA, United States), stearoyl–CoA desaturase 1 (SCD1) (ab19862 Abcam, Cambridge, UK), mevalonate kinase (MVK) (ab154515, Abcam), acetyl-CoA acetyltransferase 1 (ACAT1) (ab168342, Abcam), ACAT2 (ab131215, Abcam), glutathione peroxidase 4 (GPX4) (sc166120, Santa Cruz Biotechnology, Dallas, Texas, United States), β-Hydroxy β-methylglutaryl-CoA reductase (HMGCR) (ab174830, Abcam).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The primers involved in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The antibodies used in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: MPs induced ferroptosis in spleen. (A) Western Blot results in ferroptosis-related proteins. (B) Quantitative analysis of GPX4, FTH1, and SLC7A11 protein expression. (C) Ferroptosis-related factors of mRNA expression changes. Data were expressed as mean ± SD, n = 3. *, **, ***, denotes: p < 0.05, 0.01, and 0.001, respectively.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques: Western Blot, Expressing

Journal: Nature Communications

Article Title: IRE1α determines ferroptosis sensitivity through regulation of glutathione synthesis

doi: 10.1038/s41467-024-48330-0

Figure Lengend Snippet: GCLC Glutamate-Cysteine Ligase Catalytic Subunit, GCLM Glutamate-Cysteine Ligase Modifier Subunit, GPX4 Glutathione Peroxidase 4, GSS glutathione synthetase, LOOH lipid hydroxide, LOH alcohol, BSO buthionine sulfoximine. SLC3A2 Solute Carrier Family 3 Member 2. SLC7A11 Solute Carrier Family 7 Member 11.

Article Snippet: The following gel composition and running conditions were used for detecting (1) phospho-IRE1α: 5% Phos-tag gel with 25 μM Phos-bind acrylamide, 60 V for 30 min followed by 100 V for 3 h; (2) phospho-PERK: 5% Phos-tag gel with 3.5 μM Phos-bind acrylamide, 15 mA for 30 min followed by 5 mA for 9.5 h. 40–60 μg of protein per sample was electrophoresed and transferred onto nitrocellulose membrane (Bio-Rad) for immunoblotting using antibodies against IRE1α (3294, Cell Signaling), XBP1 (ab220783, Abcam), PERK (3192, Cell Signaling), phospho-GCN2 (ab75836, Abcam), GCN2 (3302, Cell Signaling), phospho-eIF2α (9721, Cell Signaling), eIF2α (2103, Cell Signaling), ATF4 (11815, Cell Signaling), ATF6 (CosmoBio, BAM-73-500-EX), GCLC (sc-390811, Santa Cruz Biotechnology), GCLM (sc-22754, Santa Cruz Biotechnology), GPX4 (MAB5457, R&D Systems), SLC7A11 (12691, Cell Signaling), ACSL4 (SAB2701949, Sigma Aldrich), phosphor-JNK (9251, Cell Signaling), JNK (9252, Cell Signaling), LC3B (NB600-1384, Novus), Caspase-3 (9662, Cell Signaling), luciferase (NB600-307, Novus Biologicals), calreticulin (ab2907, Abcam), HSP90 (sc-13119, Santa Cruz Biotechnology), GAPDH (8884, Cell Signaling) and β-actin (sc-1615, Santa Cruz Biotechnology) all with 1:1000 dilution in 3% (w/v) BSA (Sigma Aldrich).

Techniques:

Journal: Environmental Health and Preventive Medicine

Article Title: Chronic noise exposure induces Alzheimer’s disease-like neuropathology and cognitive impairment via ferroptosis in rat hippocampus

doi: 10.1265/ehpm.24-00126

Figure Lengend Snippet: Noise exposure induced ferroptosis in hippocampus of rats. (A–B) Changes in expression levels of GPX4, FTH1, and SLC7A11 in the rat hippocampus (n = 3). (C) Immunohistochemistry of GPX4 in rat hippocampus. (D–G) Changes in levels of MDA, SOD, GSH, and GSH-Px in the rat hippocampus. *, P < 0.05, **, P < 0.01.

Article Snippet: Paraffin sections of rat brain were deparaffinized with primary antibodies mouse GFAP (sc-166458, 1:500; Santa Crutz Biotechnology), rabbit Iba-1 (GB153502-100, 1:500; Servicebio) and rabbit GPX4 (TA376761, 1:100; ORIGENE) were incubated overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry