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D-EVs Counteract NPC Senescence by Suppressing Ferroptosis. (A) KEGG pathway analysis of DEGs in senescent NPCs following treatment with D-EVs or not. (B-C) GSEA plots showing significant enrichment of ferroptosis and cell cycle in senescent NPCs. (D-E) Heatmap quantification of key genes involved in ferroptosis and cell cycle. (F) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with different experimental conditions. (G) Representative images of C11-BODIPY 581/591 staining to detect lipid <t>peroxidation</t> (green) in the control, TBHP, Era, Era + Fer-1, or TBHP + Fer-1 groups. (H-I) Quantitative assessment of malondialdehyde <t>(MDA)</t> levels (H) and glutathione (GSH) levels (I) in the control, TBHP, N-EVs, D-EVs, or D-EVs + Era groups. (J) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with PBS, N-EVs, D-EVs, or D-EVs + Era. (K) Confocal analysis of GPX4 with IF staining in the control, TBHP, N-Evs, D-EVs, and D-EVs + Era group. (L) Flow cytometry analysis of cell cycle distribution in the above experimental conditions. Statistical comparisons were performed between the experimental group and the TBHP-induced group. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
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D-EVs Counteract NPC Senescence by Suppressing Ferroptosis. (A) KEGG pathway analysis of DEGs in senescent NPCs following treatment with D-EVs or not. (B-C) GSEA plots showing significant enrichment of ferroptosis and cell cycle in senescent NPCs. (D-E) Heatmap quantification of key genes involved in ferroptosis and cell cycle. (F) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with different experimental conditions. (G) Representative images of C11-BODIPY 581/591 staining to detect lipid <t>peroxidation</t> (green) in the control, TBHP, Era, Era + Fer-1, or TBHP + Fer-1 groups. (H-I) Quantitative assessment of malondialdehyde <t>(MDA)</t> levels (H) and glutathione (GSH) levels (I) in the control, TBHP, N-EVs, D-EVs, or D-EVs + Era groups. (J) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with PBS, N-EVs, D-EVs, or D-EVs + Era. (K) Confocal analysis of GPX4 with IF staining in the control, TBHP, N-Evs, D-EVs, and D-EVs + Era group. (L) Flow cytometry analysis of cell cycle distribution in the above experimental conditions. Statistical comparisons were performed between the experimental group and the TBHP-induced group. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
Hydrogen Peroxide Assay Kit, supplied by Shanghai Yuanye Biochemicals, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid <t>peroxidation</t> levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.
Lipid Peroxidation Assay Kit, supplied by Nanjing Jiancheng Bioengineering Research Institute Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid <t>peroxidation</t> levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.
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Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid <t>peroxidation</t> levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.
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Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid <t>peroxidation</t> levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.
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Image Search Results


D-EVs Counteract NPC Senescence by Suppressing Ferroptosis. (A) KEGG pathway analysis of DEGs in senescent NPCs following treatment with D-EVs or not. (B-C) GSEA plots showing significant enrichment of ferroptosis and cell cycle in senescent NPCs. (D-E) Heatmap quantification of key genes involved in ferroptosis and cell cycle. (F) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with different experimental conditions. (G) Representative images of C11-BODIPY 581/591 staining to detect lipid peroxidation (green) in the control, TBHP, Era, Era + Fer-1, or TBHP + Fer-1 groups. (H-I) Quantitative assessment of malondialdehyde (MDA) levels (H) and glutathione (GSH) levels (I) in the control, TBHP, N-EVs, D-EVs, or D-EVs + Era groups. (J) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with PBS, N-EVs, D-EVs, or D-EVs + Era. (K) Confocal analysis of GPX4 with IF staining in the control, TBHP, N-Evs, D-EVs, and D-EVs + Era group. (L) Flow cytometry analysis of cell cycle distribution in the above experimental conditions. Statistical comparisons were performed between the experimental group and the TBHP-induced group. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: D-EVs Counteract NPC Senescence by Suppressing Ferroptosis. (A) KEGG pathway analysis of DEGs in senescent NPCs following treatment with D-EVs or not. (B-C) GSEA plots showing significant enrichment of ferroptosis and cell cycle in senescent NPCs. (D-E) Heatmap quantification of key genes involved in ferroptosis and cell cycle. (F) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with different experimental conditions. (G) Representative images of C11-BODIPY 581/591 staining to detect lipid peroxidation (green) in the control, TBHP, Era, Era + Fer-1, or TBHP + Fer-1 groups. (H-I) Quantitative assessment of malondialdehyde (MDA) levels (H) and glutathione (GSH) levels (I) in the control, TBHP, N-EVs, D-EVs, or D-EVs + Era groups. (J) Western blot analysis of key ferroptosis (GPX4, SLC7A11, ACSL4) and senescence (p21, P16) markers in NPCs following treatment with PBS, N-EVs, D-EVs, or D-EVs + Era. (K) Confocal analysis of GPX4 with IF staining in the control, TBHP, N-Evs, D-EVs, and D-EVs + Era group. (L) Flow cytometry analysis of cell cycle distribution in the above experimental conditions. Statistical comparisons were performed between the experimental group and the TBHP-induced group. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The level of lipid peroxidation was measured by Lipid Peroxidation MDA Assay Kit (Beyotime, China) according to the manufacturer's protocol.

Techniques: Western Blot, Staining, Control, Flow Cytometry

Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid peroxidation levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.

Journal: iScience

Article Title: Ceritinib induces ferroptosis via TRIM21-mediated GLUT1 ubiquitination and AMPK-driven metabolic reprogramming in breast cancer

doi: 10.1016/j.isci.2026.115846

Figure Lengend Snippet: Ceritinib induces ferroptosis in breast cancer cells (A–D) Cell viability assays were conducted in MDA-MB-231 and MCF-7 cells treated with ceritinib alone or in combination with the ferroptosis inhibitor ferrostatin-1 (Fer-1, 15 μM), the autophagy inhibitor 3-methyladenine (3-MA, 25 μM), the apoptosis inhibitor Z-VAD-FMK (Z-VAD-FMK, 20 μM), or the necroptosis inhibitor Necrostatin-1 (Nec-1, 25 μM) ( n = 3). (E) Western blot analysis was performed to evaluate autophagy-related proteins LC3, p62, and beclin1 in response to ceritinib treatment ( n = 3). (F) Western blot was used to detect the expression of NCOA4 and FTH1 after ceritinib exposure ( n = 3). (G) Intracellular Fe 2+ concentrations were measured using a commercial iron assay kit ( n = 3). (H) Western blot analysis was carried out to assess the expression levels of GPX4, SLC7A11, FSP1, ACSL4, NRF2, KEAP1, and COX2 ( n = 3). (I and J) Lipid peroxidation levels were evaluated by measuring LPO and MDA contents in ceritinib-treated cells ( n = 3). (K) The GSH/GSSG ratio was determined to assess the cellular redox status ( n = 3). (L) NADPH levels were measured to evaluate the oxidative state of the cells treated with ceritinib ( n = 3). (M) ROS accumulation was quantified using fluorescent probes in ceritinib-treated cells ( n = 3). Significance was assessed by Student’s t test, one-way or two-way ANOVA and Tukey’s post hoc test. The data are shown as the mean ± SEM.

Article Snippet: Lipid Peroxidation Assay Kit , Nanjing Jiancheng Bioengineering Institute , Cat# A106-1-1.

Techniques: Western Blot, Expressing, Iron Assay