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: Increased mitochondrial respiration renders F12-cultured cells BSO-sensitive. ( A, B ) Oxygen consumption rate (left) in F12 or F12AA-cultured A549 (A) or H838 (B) cells treated with 0.5 μM oligomycin (Oligo), 1 μM FCCP, and 0.5 μM rotenone (Rot), as indicated. Graphs (right) showing parameter data extracted from the oxygen consumption rate and extracellular acidification rate (n = 14 in A549 cells; n = 10 in F12 and 4 in F12AA-cultured H838 cells). ( C , D ) Oxygen consumption rate (left) in F12-cultured A549 (C) or H838 (D) cells transfected with control siRNA or siRNA targeting ATF4 mRNA and assayed as in (A, B). Graphs (right) show extracted parameters (n = 15 for A549; n = 21 for ATF4 siRNA and n = 13 for control siRNA in H838). ( E ) MitoSox fluorescence images of A549 (left) and H838 (right) cells cultured in F12 or F12AA medium. Cells were treated with 30 μM BSO for 40 h (A549) or 48 h (H838). Graphs show IncuCyte-based fluorescence quantification over time. Scale bar, 100 μm. ( F ) MitoSox fluorescence images of A549 cells cultured in F12 or F12AA medium and treated for 24h with 50 μM BSO, 50 μM BSO + 20 μM mito-TEMPO (MT), or control. MitoSox was added during the last 90 min of treatment; cells were then washed and quantified by live imaging. The graph shows IncuCyte-based fluorescence quantification. Scale bar, 100 μm; n = 8–10. ( G ) Confocal microscopy images of F12-cultured A549 cells treated with 100 μM BSO for 24h showing reduced (pink) and oxidized (green) BODIPY-C11 in combination with mitotracker deep red (red). Corresponding graphs show pixel-wise colocalization (Mander's coefficient) of oxidized BODIPY-C11 and mitotracker deep red in F12-cultured A549 (left) or H838 (right) cells treated with 100 μM BSO for 24 h or controls. n = 6–9 visual fields in A549 cells and 48 visual fields in H838 cells. Scale bar, 10 μm. ( H ) Oxidized BODIPY-C11 fluorescence images of A549 cells cultured in F12 medium and treated for 12 h with 100 μM BSO, 100 nM rotenone, 100 nM oligomycin, or combinations of BSO + rotenone or BSO + oligomycin, and controls. Graph shows IncuCyte-based fluorescence quantification. Scale bar, 50 μm. ( I ) Oxidized BODIPY-C11 fluorescence images of A549 cells cultured in F12 medium and treated for 24 h with 100 μM BSO, BSO + 20 μM mito-TEMPO (MT), or control. Graph shows IncuCyte-based fluorescence quantification. Scale bar, 100 μm. ( J ) Oxidized MitoPerOx fluorescence images of A549 (left) and H838 (right) cells cultured in F12 or F12AA medium. Cells were treated with 30 μM BSO or control for 48 h. Graphs show IncuCyte-based fluorescence quantification over time. Scale bar, 100 μm. ( K) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 100 nM oligomycin, 100 nM rotenone, 20 μM mito-TEMPO + rotenone, or mito-TEMPO + oligomycin, or control for 72 h. ( L ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 100 nM oligomycin, 100 nM rotenone, 5 μM ferrostatin-1 (FER) + rotenone, 5 μM liproxstatin-1 (LIP) + rotenone, ferrostatin-1 + oligomycin, or liproxstatin-1 + oligomycin, or control for 72 h. ( M ) Dose response curves of F12-cultured A549 cells treated with BSO in combination with 0.5 μM FCCP, or control for 72 h. ( N ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 25 or 50 μM mito-TEMPO, or control for 72 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints unless otherwise indicated. Error bars show SEM. ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

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: Cell Culture, Transfection, Control, Fluorescence, Imaging, Confocal Microscopy

Journal: Redox Report : Communications in Free Radical Research

Article Title: Prophylactic quercetin administration attenuates pulmonary fibrosis via ferroptosis-resistant priming of alveolar epithelial cells

doi: 10.1080/13510002.2026.2632434

Figure Lengend Snippet: QUE-induced adaptive stress is essential for its preventive effects. (A) Schematics of QUE-, Mito-TEMPO – and Z-VAD-FMK-prevention setup. (B-D) The levels of ROS (B), GSH (C), GPx4 protein expression (D) in QUE and Miti-TEMPO co-preconditioned MLE-12 cells. (E-F) Cell viability (E) and LDH release (F) of QUE and Miti-TEMPO co-preconditioned MLE-12 cells following BLM challenge. (G-I) The levels of ROS (B), GSH (C), GPx4 protein expression (D) in QUE and Z-VAD-FMK co-preconditioned MLE-12 cells. (J-K) Cell viability (J) and LDH release (K) of QUE and Z-VAD-FMK co-preconditioned MLE-12 cells following BLM challenge. Data are presented as mean ± SEM. Statistical analysis was performed using Kruskal-Wallis H test with Dunn’s post hoc test for the data in panel C, and one-way ANOVA with Tukey’s post hoc test for the rest. *, **, and *** indicate p < 0.05, p < 0.01, p < 0.001 vs. untreated controls, respectively; #, ##, ### indicate p < 0.05, p < 0.01, p < 0.001 vs. BLM, respectively; &, &&, &&& indicate p < 0.05, p < 0.01, and p < 0.001 vs. QUE, respectively.

Article Snippet: In separate in vitro experiments, MLE-12 cells were subjected to various treatments: 100, 5 and 1 μM BLM for 24 h, 2 μM ferrostatin-1 (Fer-1; MedChemExpress, HY-100579) for 24 h, 5 μM Mito-TEMPO (MedChemExpress, HY-112879) for 24 h, 5 μM Z-VAD-FMK (MedChemExpress, HY-16658B) for 24 h, 5 mM N-acetylcysteine (NAC; MedChemExpress, HY-B0215) for 1 h, and 5 μM RAS-selective lethal 3 (RSL-3; MedChemExpress, HY-100218A) for 24 h, 100, 20 and 4 μM H 2 O 2 for 24 h.

Techniques: Expressing

Journal: Biomedical Reports

Article Title: Mitochondrial oxidative stress under hypoxia promotes gastric mucosal injury in portal hypertensive gastropathy

doi: 10.3892/br.2026.2102

Figure Lengend Snippet: ROS scavenger Mito-TEMPO attenuates gastric mucosal epithelial injury from in vivo and in vitro models. (A) H&E staining of gastric mucosal tissues and analysis of gastric mucosal injury index in the mouse model treated with Mito-TEMPO or not. Scale bar, 100 µm. n=6 per group. * P<0.05 vs. the control group; # P<0.05 vs. the PHT group without Mito-TEMPO. (B) The morphology and H&E staining of GES-1 cell under normoxic and hypoxic conditions treated with Mito-TEMPO (or not) are shown. The relative cell viability analyzed by Cell Counting Kit-8 were also determined. Scale bar, 25 µm. n=6 per group. * P<0.05 vs. the normoxia group; # P<0.05 vs. the hypoxia group without Mito-TEMPO. ROS, reactive oxygen species; H&E, hematoxylin and eosin; PHT, portal hypertension; CTRL, control.

Article Snippet: To mitigate oxidative stress, mice received intraperitoneal Mito-TEMPO (10 mg/kg solubilized in PBS; cat. no. HY-112879; MedChemExpress) every other day.

Techniques: In Vivo, In Vitro, Staining, Control, Cell Counting