Journal: Frontiers in Immunology

Article Title: In vitro study: HIF-1α-dependent glycolysis enhances NETosis in hypoxic conditions

doi: 10.3389/fimmu.2025.1583587

Figure Lengend Snippet: Time-dependent effect of hypoxia on NETosis formation. The upper part shows the expression of HIF-1α and citH3 in dHL−60 cells incubated under hypoxic conditions for various times (0–4 h) with or without PMA, assessed by Western blot analysis, with β-actin and H3 as loading controls. The lower part shows the quantification of HIF-1α and citH3 protein levels normalized to β-actin or H3, respectively. n = 6. ***p < 0.001, ****p < 0.0001, ns, not significant.

Article Snippet: The workstation was purged with gas, sealed, and placed in a conventional incubator at 37°C for 4 h. To inhibit HIF-1α expression, 10 μM LW6 (HY-13671, MedChemExpress, USA) was added to the dHL-60 cells following the initial 30-minute incubation.

Techniques: Expressing, Incubation, Western Blot

Journal: Frontiers in Immunology

Article Title: In vitro study: HIF-1α-dependent glycolysis enhances NETosis in hypoxic conditions

doi: 10.3389/fimmu.2025.1583587

Figure Lengend Snippet: LW6 inhibits HIF-1α expression and reverses hypoxia-induced NETosis. The upper part shows the expression of HIF-1α and citH3 in differentiated dHL−60 cells incubated under hypoxic conditions for 4 h with or without LW6, assessed by Western blot analysis, with β-actin and H3 as loading controls. The lower part shows the quantification of HIF-1α and citH3 protein levels normalized to β-actin or H3, respectively. n = 3 biological replicates, with three technical replicates per sample. ****p < 0.0001, ns, not significant.

Article Snippet: The workstation was purged with gas, sealed, and placed in a conventional incubator at 37°C for 4 h. To inhibit HIF-1α expression, 10 μM LW6 (HY-13671, MedChemExpress, USA) was added to the dHL-60 cells following the initial 30-minute incubation.

Techniques: Expressing, Incubation, Western Blot

Journal: Frontiers in Immunology

Article Title: In vitro study: HIF-1α-dependent glycolysis enhances NETosis in hypoxic conditions

doi: 10.3389/fimmu.2025.1583587

Figure Lengend Snippet: HIF-1α plays a key regulatory role in hypoxia-induced glycolysis and NETosis. dHL−60 were incubated under hypoxic conditions for various times (4 h) with or without LW6 (A) or Bay-876 (B) , and the expression of GLUT-1, HK2, PFK1, GAPDH, ENO1, PKM2, LDHA, and citH3 were detected by western blot. Expression of glycolysis‐related enzymes and citH3 protein levels were normalized to the β–actin or H3 protein, respectively. n = 3 biological replicates, with three technical replicates per sample. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant.

Article Snippet: The workstation was purged with gas, sealed, and placed in a conventional incubator at 37°C for 4 h. To inhibit HIF-1α expression, 10 μM LW6 (HY-13671, MedChemExpress, USA) was added to the dHL-60 cells following the initial 30-minute incubation.

Techniques: Incubation, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: In vitro study: HIF-1α-dependent glycolysis enhances NETosis in hypoxic conditions

doi: 10.3389/fimmu.2025.1583587

Figure Lengend Snippet: HIF-1α-dependent glycolysis regulates NETosis during hypoxia. (A) Representative fluorescence micrographs depicting NET-release of neutrophils. (B) Quantification of the mean fluorescence intensity (MFI) of each group of SYTOX Green. (C) Percentage of NETosis. n=6. ***p < 0.001, ****p < 0.0001.

Article Snippet: The workstation was purged with gas, sealed, and placed in a conventional incubator at 37°C for 4 h. To inhibit HIF-1α expression, 10 μM LW6 (HY-13671, MedChemExpress, USA) was added to the dHL-60 cells following the initial 30-minute incubation.

Techniques: Fluorescence

Journal: Frontiers in Immunology

Article Title: In vitro study: HIF-1α-dependent glycolysis enhances NETosis in hypoxic conditions

doi: 10.3389/fimmu.2025.1583587

Figure Lengend Snippet: Graphical Abstract. Under hypoxic conditions, HIF-1α is activated in the neutrophil nucleus. This activation promotes the transcription of key glycolytic enzymes, including GLUT1, HK2, PFK1, GAPDH, ENO1, PKM2, and LDHA, thereby enhancing intracellular glycolysis and increasing lactate production. Elevated lactate may activate PADI4, which in turn induces histone citrullination and chromatin decondensation, leading to the release of NETs into the extracellular space (left panel). Treatment with HIF-1α inhibitor LW6 or glycolysis inhibitor Bay-876 blocks this pathway, effectively reversing NETosis under hypoxic conditions (right panel), highlighting the critical role of HIF-1α-dependent glycolysis in NETosis under low oxygen stress.

Article Snippet: The workstation was purged with gas, sealed, and placed in a conventional incubator at 37°C for 4 h. To inhibit HIF-1α expression, 10 μM LW6 (HY-13671, MedChemExpress, USA) was added to the dHL-60 cells following the initial 30-minute incubation.

Techniques: Activation Assay

Journal: Molecular Therapy

Article Title: Perivascular Stem Cell-Derived Cyclophilin A Improves Uterine Environment with Asherman’s Syndrome via HIF1α-Dependent Angiogenesis

doi: 10.1016/j.ymthe.2020.05.015

Figure Lengend Snippet: hPVSCs Facilitate HIF1α-Dependent Angiogenesis to Improve Fertility in Mice with AS

Article Snippet: Pharmacological Modulators of HIF1α Echinomycin (EC; HIF1α inhibitor, 1 mg/kg, Bioviotica, Liestal, Switzerland) or dimethyloxalylglycine (DMOG; PHD inhibitor; HIF1α stabilizer; 8 mg, Enzo Life Sciences, Lausen, Switzerland) was used to either inhibit or stabilize HIF1α activity in mice with AS, respectively.

Techniques:

Journal: Nature Communications

Article Title: Treg-γδ T cell axis determines sexual dimorphism in hepatocarcinogenesis

doi: 10.1038/s41467-026-69603-w

Figure Lengend Snippet: Umap projection visualizing total ( a ) and gender-specific ( b ) of T cell clusters of human HCC scRNA-seq data (male, n = 16; female, n = 15). Umap plot showing the enrichment of representative cytotoxic ( c ) and exhausted genes ( d ) (male, n = 16; female, n = 15). e Single-cell transcription levels of representative genes in CD8 + T cells and γδ T cells between male and female (male, n = 16; female, n = 15). Single-cell transcription levels of S100A4 in T cell subsets ( f ) and Tregs ( g ). h Volcano plots of DEGs (male/female) in Tregs from scRNA-seq data. The red dots indicate upregulated genes. The blue dots indicate downregulated genes (male, n = 16; female, n = 15). i The relative mRNA expression of S100A4 in human HCC samples measured by qPCR (male, n = 20; female, n = 20). j The correlation analysis between the relative mRNA expression of S100A4 or FOXP3 in human HCC samples ( n = 37). k The overall survival of Foxp3 high expression ( n = 188) and Foxp3 low expression patients ( n = 194) from UCSC database. l Immunofluorescent staining of Foxp3, S100a4 and DAPI in orthotopic HCC. Scale bars, 50 μm (left). Quantification of Foxp3 + S100a4 + cells (right) ( n = 8). Mouse splenic Tregs were cultured in normoxia or hypoxia for 48 h. The relative mRNA expression of S100a4 by qPCR ( m ). The western blotting of Hif1α and S100a4 protein ( n ) ( n = 3). o Cut&Tag analysis was used to assess the interaction between Hif1α and S100a4 in Tregs. The relative expression of S100a4 with specific primers targeting the sequences located at +76 to +195 of S100a4 promoter locus by qPCR ( n = 4). Data represent three independent experiments. Data are shown as the mean ± SEM. P value of Pearson r correlation ( j ) or Log-rank (Mantel–Cox) test ( k ) or two-tailed unpaired Student’s t test ( i , l , m and o ). For additional details, see also Supplementary Fig. and Supplementary Data . Source data are provided as a Source Data file.

Article Snippet: For experiments with the Hif1α inhibitor PX-478 (Shanghai Yuanye Bio-Technology Co., Ltd), mice were treated by intraperitoneal injection at a dose of 30 mg/kg body weight twice a week after orthotopic implantation.

Techniques: Single Cell, Expressing, Staining, Cell Culture, Western Blot, Two Tailed Test