ferroptosis inhibitor fer 1  (MedChemExpress)

Journal: Redox Biology

Article Title: A spinal USP7-Bach1 positive feedback loop drives NOX4-mediated ferroptosis in neuropathic pain

doi: 10.1016/j.redox.2026.104153

Figure Lengend Snippet: FER-1 attenuates neuropathic pain and spinal ferroptosis in the SNI model. (A) Mechanical and thermal hyperalgesia were evaluated by the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) in Sham + Vehicle, SNI + Vehicle, and SNI + FER-1 groups. Behavioral tests were conducted at day 0 and on days 6, 7, 8, 9, 10, and 11 post-surgery (n = 9 per group). (B) Representative Western blot images and quantitative analysis of 4-HNE in spinal cord lysates 11 days post-SNI (n = 6 per group). (C) Representative double immunofluorescence images showing co-localization of 4-HNE (red) with the neuronal marker NeuN (green) in the spinal dorsal horn. Scale bar: 100 μm. White boxes indicate higher magnification views of representative neurons (n = 3 per group). (D-G) Representative Western blot images and quantitative analysis of ferroptosis-related proteins (ACSL4, SLC7A11 and GPX4) in the spinal cord. β-actin served as the loading control (n = 6 per group). (H–I) Biochemical quantification of Fe 2+ and MDA levels in spinal cord tissues (n = 6 per group). (J) Representative transmission electron microscopy (TEM) images and quantitative analysis of mitochondrial morphology in spinal neurons. Red arrowheads indicate classical shrunken mitochondria with increased electron density of the bilayer membrane, accompanied by decreased mitochondrial cristae and mitochondrial membrane rupture; blue arrowheads indicate reduced or vanishing cristae; and black arrowheads indicate outer mitochondrial membrane rupture. Scale bar: 500 nm (n = 3 per group). Significance was determined using one-way ANOVA or two-way ANOVA followed by Bonferroni's post-hoc tests. Data are expressed as mean ± SEM. (∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 vs. Sham + Vehicle group; # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 vs. SNI + Vehicle group). The value of n represents the number of independent biological samples. All molecular experiments were independently repeated at least three times with consistent results.

Article Snippet: In in vivo experiments, the ferroptosis inhibitor FER-1 (10 mg/kg, i.p., Hy-100579) [ ], the Bach1 inhibitor Bach1-IN-1 (also known as HPPE, 10 mg/kg, i.g., HY-153040 , MCE, China), the NOX4 inhibitor GLX351322 (5 mg/kg, i.p., HY-100111, MCE, China), and the USP7 inhibitor P005091 (15 mg/kg, i.p., HY-15667 , MCE, China) were administered once daily for 5 consecutive days.

Techniques: Western Blot, Immunofluorescence, Marker, Control, Transmission Assay, Electron Microscopy, Membrane

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Copper deprivation induced by SLC31A1 knockdown attenuates ferroptosis. ( A ) Cell viability of AsPC-1 cells treated with different doses of RSL3, ML162, ML210, or erastin in NC and SLC31A1 knockdown AsPC-1 cells. ( B ) Cell viability of AsPC-1 cells treated with different doses of CDDP, staurosporine, paclitaxel, bortezomib, JTC-801, or elesclomol-Cu in NC and SLC31A1 knockdown AsPC-1 cells. ( C-E ) Propidium iodide (PI) staining of NC and SLC31A1 knockdown AsPC-1 (C), MiaPaCa-2 (D), and CFPAC-1 (E) cells treated with RSL3 at the indicated concentrations in the presence or absence of ferroptosis inhibitors (ferrostatin-1/Fer-1, 5 μM or desferrioxamine/DFO, 20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( F ) Lipid peroxidation of NC and SLC31A1 knockdown AsPC-1 cells treated with RSL3 (0.25 μM) for 2 h. (G) Transmission electron microscopy of NC and SLC31A1 knockdown AsPC-1 cells treated with RSL3 (2.5 μM). Scale bar: 2 μm or 500 nm. ( H ) Confocal imaging of Cu-probe and Mito-tracker in the NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1. Hoechst (blue) is used as a nuclear counterstain. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 2 μm. ( I ) Western blot analysis of lysates from NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1. ( J ) Cell viability of NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1, followed by treatment with different doses of RSL3 or erastin. ( K ) PI staining of NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1, followed by treatment with RSL3 at the indicated concentrations in the presence or absence of Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( L ) Western blot analysis of lysates from SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or SLC31A1 WT or SLC31A1 M154A plasmid. ( M ) PI staining of SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or SLC31A1 WT or SLC31A1 M154A plasmid, followed by treatment with RSL3 at the indicated concentrations in the presence or absence of Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test.

Article Snippet: Ferrostatin-1 (Fer-1) , Selleck Chemicals , S7243.

Techniques: Knockdown, Staining, Transmission Assay, Electron Microscopy, Imaging, shRNA, Transfection, Plasmid Preparation, Western Blot

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Pharmacological induction of copper deprivation upregulates SLC7A11 and suppresses ferroptosis. ( A ) Western blot analysis of lysates from AsPC-1 cells treated with the indicated concentrations of TM for 24 h. ( B ) SLC7A11 mRNA level in AsPC-1 cells treated with TM at the indicated concentrations for 24 h. ( C ) Western blot analysis of lysates from AsPC-1 cells treated with TEPA at the indicated concentrations for 24 h. ( D ) SLC7A11 mRNA level in AsPC-1 cells treated with TEPA at the indicated concentrations for 24 h. ( E ) GSH level in the AsPC-1 cells treated with TM (50 μM) or TEPA (1 mM) for 24 h. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( F ) Cystine uptake level in the AsPC-1 cells treated with TM (50 μM) or TEPA (1 mM) for 24 h. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( G ) Cell viability of PDAC cells treated with RSL3 in the presence or absence of TM (50 μM) or TEPA (1 mM) for 24 h. ( H-J ) Propidium iodide (PI) staining of AsPC-1 (H), MiaPaCa-2 (I), and CFPAC-1 (J) cells treated with RSL3 at the indicated concentrations in the presence or absence of TM (50 μM) or TEPA (1 mM) with or without Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( K ) Lipid peroxidation of AsPC-1 cells treated with RSL3 (0.25 μM) in the presence or absence of TM (50 μM) or TEPA (1 mM) for 2 h. ( L ) Cell viability of KYSE-410, NCI–H1666, SW579, or OS-RC-2 cells treated with RSL3 at the indicated concentrations in the presence or absence of TM (50 μM) for 12 h.

Article Snippet: Ferrostatin-1 (Fer-1) , Selleck Chemicals , S7243.

Techniques: Western Blot, Staining

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: SLC7A11 is essential for copper deprivation-mediated ferroptosis resistance. ( A ) NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC), SLC7A11 siRNA (siSLC7A11). Western blot analysis of lysates from the indicated AsPC-1 cells. ( B ) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). Western blot analysis of lysates from the indicated AsPC-1 cells treated with or without TM (50 μM) for 24 h. ( C ) NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). Propidium iodide (PI) staining of the indicated AsPC-1 cells treated with RSL3 at the indicated concentrations in the presence or absence of Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( D ) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). PI staining of the indicated AsPC-1 cells treated with RSL3 at the indicated concentrations in the presence or absence of TM (50 μM) with or without Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( E ) NC and SLC31A1 knockdown AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). Lipid peroxidation of the AsPC-1 cells treated with RSL3 (0.25 μM) for 2 h. ( F ) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). Lipid peroxidation of the indicated AsPC-1 cells treated with RSL3 (0.25 μM) in the presence or absence of TM (50 μM) for 2 h. ( G ) PI staining of the NC and SLC31A1 knockdown AsPC-1 cells treated with RSL3 at the indicated concentrations in the presence or absence of BSO (500 μM) with or without Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( H ) PI staining images of the indicated AsPC-1 cells treated with or without BSO (500 μM) plus TM sulfate (50 μM) with RSL3 at the indicated concentrations with or without Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test.

Article Snippet: Ferrostatin-1 (Fer-1) , Selleck Chemicals , S7243.

Techniques: Knockdown, shRNA, Transfection, Western Blot, Staining

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: SLC7A11 is essential for copper deprivation-mediated ferroptosis resistance. ( A ) Vector or flag-SLC7A11 overexpressed AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11). Western blot analysis of lysates from the indicated AsPC-1 cells. ( B ) AsPC-1 cells were transfected with a vector or a plasmid encoding Flag-SLC7A11 and then treated with TM (50 μM) for 24h. Western blot analysis of lysates from the indicated AsPC-1 cells. ( C ) NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11) with or without transfection with a blank vector or a vector encoding SLC7A11. PI staining of indicated AsPC-1 cells treated with RSL3 (0.25 μM) in the presence or absence of Fer-1 (2.5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 50 μm. ( D ) AsPC-1 cells were transfected with non-targeting scrambled siRNA (siNC) or SLC7A11 siRNA (siSLC7A11) with or without transfection with a blank vector or a vector encoding SLC7A11. PI staining images of indicated AsPC-1 cells treated with RSL3 (0.25 μM) or TM (50 μM) in the presence or absence of Fer-1 (2.5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 50 μm.

Article Snippet: Ferrostatin-1 (Fer-1) , Selleck Chemicals , S7243.

Techniques: Plasmid Preparation, Transfection, Western Blot, Knockdown, shRNA, Staining

Journal: Cells

Article Title: Key Molecular Events in PM 2.5 -Induced Lung Injury: Autophagy and Ferroptosis Mediated by the miR-212-5p / RASSF1 Axis

doi: 10.3390/cells15090823

Figure Lengend Snippet: PM 2.5 decreases the activity of RLE-6TN cells and induces cellular oxidative stress. ( A ) CCK-8 assay for cell viability. ( B ) ROS levels in PM 2.5 -exposed RLE-6TN cells (scale bar = 300 µm). ( C ) Changes in intracellular malondialdehyde (MDA) levels following PM 2.5 stimulation of RLE-6TN cells. ( D ) The effects of PM 2.5 stimulation on cellular superoxide dismutase (SOD) activity. ( E ) Changes in intracellular Fe 2+ levels following treatment with different concentrations of PM 2.5 alone or combined with Fer-1 pretreatment. ( F ) Changes in intracellular Fe 2+ levels after PM 2.5 intervention for different time points with or without Fer-1 pretreatment. Data are mean ± SD. Significance: ns ( p > 0.05), * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control.

Article Snippet: In these experiments, cells were pretreated with the ferroptosis-specific inhibitor Fer-1 (MedChemExpress, Monmouth Junction, NJ, USA) for 1 h, and changes in the expression of ferroptosis-related markers were subsequently detected to assess the extent to which Fer-1 rescued ferroptosis.

Techniques: Activity Assay, CCK-8 Assay, Control

Journal: Cells

Article Title: Key Molecular Events in PM 2.5 -Induced Lung Injury: Autophagy and Ferroptosis Mediated by the miR-212-5p / RASSF1 Axis

doi: 10.3390/cells15090823

Figure Lengend Snippet: PM 2.5 induces autophagy and ferroptosis in RLE-6TN cells. ( A , B ) Expression levels of LC3-II/LC3-I and p62 were detected by Western blotting. ( C – F ) Western blotting was used to detect the expression levels of the ferroptosis-related proteins ACSL4 and GPX4. Two groups were formed: one exposed to PM 2.5 alone and one pretreated with ferritin (Fer-1) and then exposed to PM2.5. Data are presented as mean ± SD. Significance: ns = not significant ( p > 0.05), p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) vs. control group.

Article Snippet: In these experiments, cells were pretreated with the ferroptosis-specific inhibitor Fer-1 (MedChemExpress, Monmouth Junction, NJ, USA) for 1 h, and changes in the expression of ferroptosis-related markers were subsequently detected to assess the extent to which Fer-1 rescued ferroptosis.

Techniques: Expressing, Western Blot, Control

Journal: Cells

Article Title: Key Molecular Events in PM 2.5 -Induced Lung Injury: Autophagy and Ferroptosis Mediated by the miR-212-5p / RASSF1 Axis

doi: 10.3390/cells15090823

Figure Lengend Snippet: miR-212-5p promotes cellular oxidative stress, autophagy, and ferroptosis. ( A ) RT-qPCR of miR-212-5p in RLE-6TN cells treated with 180 ug/mL PM 2.5 . ( B ) Intracellular ROS, MDA, and SOD levels in PM 2.5 -treated RLE-6TN cells transfected with miR-212-5p mimics or inhibitor (scale bar = 300 µm). ( C , D ) Expression of the autophagy-related proteins LC3-II/LC3-I and p62 in RLE-6TN cells stimulated with 180 ug/mL PM 2.5 , as detected by Western blotting. ( E , F ) Expression of ferroptosis-related proteins ACSL4 and GPX4 in RLE-6TN cells stimulated with 180 ug/mL PM 2.5 was detected by Western blotting. ( G ) Fe 2+ levels were measured in cells from the miR-212-5p mimic and inhibitor groups, respectively. ( H ) The expression levels of ferroptosis-related proteins ACSL4 and GPX4, as well as Fe 2+ content, were assessed by Western blotting following Fer-1 pretreatment. Data are mean ± SD. Significance: ns ( p > 0.05), * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control.

Article Snippet: In these experiments, cells were pretreated with the ferroptosis-specific inhibitor Fer-1 (MedChemExpress, Monmouth Junction, NJ, USA) for 1 h, and changes in the expression of ferroptosis-related markers were subsequently detected to assess the extent to which Fer-1 rescued ferroptosis.

Techniques: Quantitative RT-PCR, Transfection, Expressing, Western Blot, Control

Journal: Cells

Article Title: Key Molecular Events in PM 2.5 -Induced Lung Injury: Autophagy and Ferroptosis Mediated by the miR-212-5p / RASSF1 Axis

doi: 10.3390/cells15090823

Figure Lengend Snippet: RASSF1 reverses the effects of PM 2.5 on cellular oxidative stress, autophagy, and ferroptosis via the PI3K/AKT/mTOR pathway. ( A ) RASSF1 expression in PM 2.5 -stimulated cells. ( B ) Validation of RASSF1 overexpression (RT-qPCR, WB, fluorescence; OE- RASSF1 -mCherry; scale bar = 300 µm). ( C – I ) Assays in cells treated with 180 ug/mL PM 2.5 (24 h) and transfected with OE- RASSF1 : ( C ) Oxidative stress markers (ROS, MDA, SOD); ( D ) Autophagy-related proteins (LC3-II/LC3-I, p62); ( E ) Intracellular Fe 2+ levels; ( F ) Ferroptosis-related proteins (ACSL4, GPX4); ( G , H ) Expression levels of the ferroptosis-related proteins ACSL4 and GPX4, as well as Fe 2+ content, following Fer-1 pretreatment; ( I ) PI3K/AKT/mTOR pathway key proteins. Data: mean ± SD. Significance: ns ( p > 0.05), * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control.

Article Snippet: In these experiments, cells were pretreated with the ferroptosis-specific inhibitor Fer-1 (MedChemExpress, Monmouth Junction, NJ, USA) for 1 h, and changes in the expression of ferroptosis-related markers were subsequently detected to assess the extent to which Fer-1 rescued ferroptosis.

Techniques: Expressing, Biomarker Discovery, Over Expression, Quantitative RT-PCR, Fluorescence, Transfection, Control

Journal: Cells

Article Title: Key Molecular Events in PM 2.5 -Induced Lung Injury: Autophagy and Ferroptosis Mediated by the miR-212-5p / RASSF1 Axis

doi: 10.3390/cells15090823

Figure Lengend Snippet: The miR-212-5p / RASSF1 axis controls autophagy and ferroptosis in cells by regulating the PI3K/AKT/mTOR signaling pathway. ( A , B ). RT-qPCR ( A ) and Western blot ( B ) analyses were performed to investigate the expression of RASSF1 , regulated by miR-212-5p , in RASSF1 -overexpressing cells treated with 180 ug/mL PM 2.5 . ( C ) Shows the expression of ferroptosis-related proteins (ACSL4 and GPX4) in RASSF1 -overexpressing cells 24 h after treatment with 180 μg/mL PM 2.5 , followed by transfection with miR-212-5p mimics or inhibitors. ( D ) Expression levels of ACSL4 and GPX4 proteins and Fe 2+ content following Fer-1 pretreatment under the same conditions as in ( C ). ( E ) Western blot analysis of key proteins in the PI3K/AKT/mTOR pathway under the same conditions as in ( C ). ( F ) Expression levels of ferroptosis-related proteins ACSL4 and GPX4, as well as Fe 2+ content, were detected by Western blot following Fer-1 rescue treatment. Data: mean ± SD. Significance: ns ( p > 0.05), * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control; # p < 0.05, ## p < 0.01 vs. control.

Article Snippet: In these experiments, cells were pretreated with the ferroptosis-specific inhibitor Fer-1 (MedChemExpress, Monmouth Junction, NJ, USA) for 1 h, and changes in the expression of ferroptosis-related markers were subsequently detected to assess the extent to which Fer-1 rescued ferroptosis.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transfection, Control