Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: F12 medium sensitizes lung cancer cells to iron-dependent lipid peroxidation . ( A, B ) Quantification by flow cytometry of oxidized BODIPY-C11 fluorescence in A549 (A) or H838 (B) cells that were cultured in RPMI or F12 medium and treated with 100 μM BSO or vehicle for 24 h. ( C ) Dose response curves for F12-cultured A549 and H838 cells treated with BSO in combination with 5 μM liproxstatin-1 (LIP-1), 5 μM ferrostatin-1 (FER-1) or 50 μM α-tocopherol (α-toco) for 72 h. ( D ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM deferoxamine (DFO) for A549 cells and 9 μM for H838 cells for 72 h. ( E ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM necrostatin-1 (NEC-1) or 10 μM ZVAD-FMK (ZVAD) for 72 h. ( F ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 4 μg/mL certolizumab (CER), 15 μM CU-CPT4a (C4a), 3 μM resatorvid (RST), or 50 μM necrostatin-1 (NEC-1) for 72 h (ND, not done). Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗∗∗P < 0.0001.

Article Snippet: Drugs used were auranofin (A6733; Sigma-Aldrich), α-tocopherol (T3251; Sigma-Aldrich), bafilomycin A1 (SML1661; Sigma-Aldrich), certolizumab pegol (HYP9953; MedChemExpress), chloroquine (C6628; Sigma-Aldrich), CU-CPT4A (HY-108473; MedChemExpress), deferoxamine (D9533; Sigma-Aldrich), erastin (e7781; Sigma Aldrich), FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) (HY-100410; MedChemExpress), ferrostatin-1 (SML0583; Sigma-Aldrich), l -buthionine-sulfoximine (b2515; Sigma-Aldrich), mito-TEMPO (HY–W001187; MedChemExpress), necrostatin-1 (N9037; Sigma-Aldrich), liproxstatin-1 (SML1414; Sigma-Aldrich), oligomycin (HY–N6782; MedChemExpress), puromycin dihydrochloride (A1113803; Gibco), rotenone (HY–B1756; MedChemExpress), RSL3 (SML2234; Sigma-Aldrich), resatorvid (HY-11109; MedChemExpress), and Z-VAD-FMK (V116; Sigma-Aldrich).

Techniques: Flow Cytometry, Fluorescence, Cell Culture

Journal: International Journal of Oral Science

Article Title: Cuproptosis promotes inflammatory osteolysis via GYS1-mediated glycogen metabolism

doi: 10.1038/s41368-025-00408-1

Figure Lengend Snippet: Glycogen metabolism-PPP regulates macrophages phenotype in cuproptosis. a Macrophages were treated with ES-Cu (150 nmol/L, 1:1) for 12 h after 1 μmol/L MZ-101 pre-treatment for 30 min. The level of glycogen was detected in MZ-101 treated macrophages. NADPH/NADP + ratio ( b ), GSH/GSSG ratio ( c ) and ROS level ( d ) were analyzed. Macrophages were pretreated with GYS1 siRNA and then treated with ES-Cu (150 nmol/L, 1:1) for 12 h. The glycogen level ( e ), NADPH/NADP + ratio ( f ), GSH/GSSG ratio ( g ) and ROS level ( h ) were analyzed. Macrophages were treated with ES-Cu (150 nmol/L, 1:1) after overexpressing GYS1. The glycogen level ( i ), NADPH/NADP + ratio ( j ), GSH/GSSG ratio ( k ) and ROS level ( l ) were analyzed. Data are from three independent experiments. All error bars are mean ± SEM. P values were calculated by 1-way ANOVA followed by Tukey’s multiple-comparison test

Article Snippet: ES-Cu (40 μmol/kg, 1:1; Elesclomol, MedChemExpress, HY-12040; CuCl 2 , Aladdin, C106775) was injected into the dental pulp cavity.

Techniques: Comparison

Journal: International Journal of Oral Science

Article Title: Cuproptosis promotes inflammatory osteolysis via GYS1-mediated glycogen metabolism

doi: 10.1038/s41368-025-00408-1

Figure Lengend Snippet: Glycogen metabolism regulates osteoclastogenesis in cuproptosis. a The workflow of in vitro osteoclastogenesis assay for conditioned media collection. b Osteoclasts were induced in conditioned media and then stained for TRAP-positive cells. Scale bar: 100 μm. c The quantification of TRAP-stained polynucleated (≥5 nuclei) osteoclasts that were stimulated with ES-Cu and MZ-101. d The mRNA levels of Acp5 , Oscar , Dcstamp and Fos in BMDMs treated with ES-Cu (150 nmol/L, 1:1) after 1 μmol/L MZ-101 pretreatment. e Western blot analysis of p-Src and MMP9 expression in macrophages after ES-Cu and MZ-101 treatment. Data are from 3 independent experiments. All error bars are mean ± SEM. P values were calculated by 1-way ANOVA followed by Tukey’s multiple-comparison test

Article Snippet: ES-Cu (40 μmol/kg, 1:1; Elesclomol, MedChemExpress, HY-12040; CuCl 2 , Aladdin, C106775) was injected into the dental pulp cavity.

Techniques: In Vitro, Staining, Western Blot, Expressing, Comparison

Journal: International Journal of Oral Science

Article Title: Cuproptosis promotes inflammatory osteolysis via GYS1-mediated glycogen metabolism

doi: 10.1038/s41368-025-00408-1

Figure Lengend Snippet: Copper binding to H3K27me3 epigenetically suppresses GYS1 expression. a Schematic diagram illustrating the copper binding with H3K27me3. b Western blot analysis of H3K27me3 expression in ES-Cu treated macrophages. c The co-localization of copper (red) and H3K27me3 (green) was detected by confocal. Scale bar: 5 μm. d ChIP-qPCR for H3K27me3 at the GYS1 promoters in macrophages. e Macrophages were pretreated with 1 μmol/L GSK-4J for 30 min, then treated with ES-Cu (150 nmol/L, 1:1) for 12 h. Western blot analysis of H3K27me3 and GYS1 expression in macrophages. f Cell viability of macrophages was analyzed. g The osteoclasts were induced and the representative images of TRAP staining were shown. Scale bar: 100 μm. h The quantification of TRAP-stained polynucleated (≥5 nuclei) osteoclasts. Data are from 3 independent experiments. All error bars are mean ± SEM. P values were calculated by 1-way ANOVA followed by Tukey’s multiple-comparison test

Article Snippet: ES-Cu (40 μmol/kg, 1:1; Elesclomol, MedChemExpress, HY-12040; CuCl 2 , Aladdin, C106775) was injected into the dental pulp cavity.

Techniques: Binding Assay, Expressing, Western Blot, ChIP-qPCR, Staining, Comparison

Journal: International Journal of Oral Science

Article Title: Cuproptosis promotes inflammatory osteolysis via GYS1-mediated glycogen metabolism

doi: 10.1038/s41368-025-00408-1

Figure Lengend Snippet: The disruption of glycogen metabolism intensifies cuproptosis and inflammatory bone loss in vivo. a Schematic illustration of the mouse calvarial osteolysis experimental procedure. ES-Cu (30 μmol/kg, 1:1) was administered daily to the calvaria of mice for 7 days, and mice were randomly assigned to receive TTM (4 mmol/kg) or MZ-101 (0.2 mmol/kg) via intraperitoneal injection every other day. b Representative micro-computed tomography (micro-CT) images of calvarial bones. Scale bars: 500 μm ( n = 6). c Representative images of H&E, TRAP and anti-DLAT body staining at calvarial bones. Scale bars of H&E and TRAP: 100 μm. Scale bar of DLAT: 50 μm ( n = 6)

Article Snippet: ES-Cu (40 μmol/kg, 1:1; Elesclomol, MedChemExpress, HY-12040; CuCl 2 , Aladdin, C106775) was injected into the dental pulp cavity.

Techniques: Disruption, In Vivo, Injection, Micro-CT, Staining