Journal: Genes & Diseases

Article Title: Non-canonical role of “S6K1–SGK1” pathway in neuronal necroptosis following traumatic brain injury

doi: 10.1016/j.gendis.2025.101876

Figure Lengend Snippet: TSZ induces necroptosis in HT22 cells. (A) LDH assay indicated an increased percentage of cell death following TSZ application, which was reduced by Nec-1 pretreatment before TSZ application. (B) PI staining (red) indicated the increased changes of necrotic cells after TSZ application and the decreased changes of necrotic cells after pretreatment with Nec-1 before TSZ application. Nuclei were stained with DAPI (blue). Scale bar: 50 μm. (C) Statistical results of the PI-positive necrotic cells. (D) Western blotting showed the increased changes of p-MLKL/MLKL and p-RIP3/RIP3 expression in HT22 cells after TSZ application or pretreatment with Nec-1 before TSZ application. (E, F) Statistical results of Western blotting. (G) Representative transmission electron microscopic images showed necroptosis after TSZ treatment. The black arrowhead indicates plasma membrane rupture. The triangle indicates mitochondrial swelling in TSZ-treated HT22 cells. Scale bar: 10 μm. n = 3 or 9. ∗ P < 0.05, ∗∗P < 0.01, and ∗∗∗∗P < 0.0001 versus the CTL group. ## P < 0.01 and #### P < 0.0001 versus the TSZ 3 h group. Two-tailed unpaired Student's t -test was used for Nec-1 comparison.

Article Snippet: Nec-1 (20 μM), PF4708671 (20 μM), and GSK650394 (15 μM) were all from MedChemExpress (New Jersey, USA) and were added 30 min before TSZ injury.

Techniques: Lactate Dehydrogenase Assay, Staining, Western Blot, Expressing, Transmission Assay, Clinical Proteomics, Membrane, Two Tailed Test, Comparison

Journal: Redox Biology

Article Title: USP20 governs tyrosine kinase inhibitors resistance through ferroptosis evasion by targeting GPX4 in cancers

doi: 10.1016/j.redox.2026.104086

Figure Lengend Snippet: TKI-resistant cancer cells exhibit reduced ferroptosis susceptibility due to increased GPX4 stability. ( A ) Development of sorafenib-resistant (Sora-R) cell models. Renal carcinoma 769-P cells were subjected to chronic sorafenib exposure (initial 0.5 μM for 5 days, followed by 1 μM dose escalation every 5 days until reaching 10 μM, then cultured in medium with 10 μM sorafenib for 3 month). Surviving populations were designated as Sora-R. The validity of this treatment was assessed in parental and Sora-R cells after 24 h sorafenib treatment using MTT assays. ( B ) The validity of sorafenib-resistant cells with colony formation assay. Sorafenib-resistant and parental cells was cultured for 7 days while treated with indicated concentration of sorafenib. The colony number was measured with Image J. ( C-D ) Parental and Sora-R 769-P cells were treated with 30 μM sorafenib for 6 h after pretreated with Tempol (50 μM), EUK-143 (50 μM), DFO (50 μM), and Fer-1 (1 μM) for 2 h, followed by quantification of total ROS using fluorescence microscope with DCFH-DA probe. Scale bar: 50 μm. ( E ) ROS generation and clearance. ( F ) Parental and Sora-R 769-P cells were treated with 30 μM sorafenib for 6 h, then quantified of GPX4 activity. ( G-H ) Parental and Sora-R 769-P cells were treated with 30 μM sorafenib for 6 h after pretreated with Tempol (50 μM), EUK-143 (50 μM), DFO (50 μM), and Fer-1 (1 μM) for 2 h, followed by quantification of lipid ROS using fluorescence microscope with C11-BODIPY 581/591 probe. Scale bar: 50 μm. ( I ) Parental and Sora-R 769-P cells were treated with 30 μM sorafenib for 6 h after pretreated with DFO (50 μM) and Fer-1 (1 μM) for 2 h, followed by quantification of MDA via ELISA kits. ( J-K ) Parental and Sora-R cells were treated with 30 μM IKE for 6 h, followed by quantification of lipid ROS using fluorescence microscope with C11-BODIPY 581/591 probe. Scale bar: 50 μm. ( L ) 769-P cells were treated with 20 μM sorafenib for 24 h after pretreated with 1 μM Fer-1, 2 μM Lip-1, 50 μM DFO, 10 μM Nec-1, 25 μM Z-VAD-FMK, 25 μM 3-MA, 50 μM CQ, and 10 μM TTM, cell viability was measured via MTT assay. ( M-N ) Immunoblot analysis of key antioxidant and ferroptosis-related proteins in parental and Sora-R cell lines. For A-D and F-L , data are presented as mean ± s.d. of n = 3 biological replicates.

Article Snippet: T2490), Afatinib (cat. no. T21312 ), GSK2643943A (cat. no. T11485 ), Tempol (cat. no. T6699), EUK-134(cat. no. T6495), DFO (cat. no. T18971 ), Nec-1 (cat. no. T1847), Z-VAD-FMK (cat. no. T 7020), 3-MA (cat. no. T1879), TTM (cat. no. T77774 ), Lip-1 (cat. no. T2376), and MTT (cat. no. T19029 ) were purchased from TargetMol.

Techniques: Cell Culture, Colony Assay, Concentration Assay, Fluorescence, Microscopy, Activity Assay, Enzyme-linked Immunosorbent Assay, MTT Assay, Western Blot

Journal: Frontiers in Microbiology

Article Title: Ferrous sulfate induces ferroptosis-like cell death in Trichosporon asahii

doi: 10.3389/fmicb.2026.1789479

Figure Lengend Snippet: Mechanisms of FeSO 4 -induced killing in Trichosporon asahii . (A) Cell survival rates after treatment with FeSO 4 (2 mM, 12 h) combined with different cell death pathway inhibitors. Inhibitor concentrations: ferroptosis inhibitor Fer-1 (20 μM), apoptosis inhibitor Z-VAD-FMK (20 μM), necroptosis inhibitor Nec-1 (50 μM), and cell death inhibitor CHO (20 μM). (B) Cell survival rates after treatment with FeSO 4 (2 mM, 12 h) combined with ferroptosis-related inhibitors. Inhibitor concentrations: Fer-1 (20 μM), Lip-1 (10 μM), NAC (5 mM), GSH (5 mM), and Dipy (100 μM). (C) Relative intracellular Fe 2+ levels (ferrous ion fluorescent probe, Ex/Em = 542/580 nm). (D) Relative intracellular ROS levels (DCFH-DA probe, Ex/Em = 488/525 nm). (E) Representative fluorescence microscopy images of ROS staining (same as D ). (F) Lipid peroxidation level (C11-BODIPY 581/591 probe, Ex/Em = 488/590 nm). (G) Malondialdehyde (MDA) content (μM). (H) JC-1 red/green fluorescence intensity ratio (Ex/Em = 535/590 nm and 485/530 nm). (I) Representative fluorescence microscopy images of JC-1 staining (red: aggregated form, high ΔΨm; green: monomeric form, low ΔΨm). (J) Relative Nile Red fluorescence intensity (Ex/Em = 485/590 nm). (K) Representative fluorescence microscopy images of Nile Red staining (neutral lipids). All quantitative data are presented as mean ± SD from three independent experiments ( n = 3). FeSO 4 was used at 2 mM for 12 h (A,B) or 24 h (C–J) . Statistical significance was determined by one-way ANOVA with Dunnett’s or Tukey’s post-hoc tests, with significance indicated as * p < 0.05, ** p < 0.01, *** p < 0.001 (compared to the control group or as specified).

Article Snippet: The ferroptosis inhibitors Ferrostatin-1 (Fer-1, 20 μM) and Liproxstatin-1 (Lip-1, 10 μM), the reactive oxygen species scavenger N-acetylcysteine (NAC, 5 mM), reduced glutathione (GSH, 5 mM), the iron chelator 2,2′-bipyridyl (Dipy, 100 μM), the apoptosis inhibitor Z-VAD-FMK (20 μM), the cell death inhibitor cycloheximide (CHO, 20 μM) and the necroptosis inhibitor Nec-1 (50 μM) were all purchased from MedChemExpress (United States).

Techniques: Fluorescence, Microscopy, Staining, Control