Journal: Nature Communications

Article Title: Hexokinase 2-mediated histone H3K18la promotes PAI-1-dependent thrombosis in acute myeloid leukemia via tumor-endothelial crosstalk

doi: 10.1038/s41467-025-65259-0

Figure Lengend Snippet: a NB4 and MOLM13 cells were exposed to 20 mM lactate for 12 or 24 hours, the levels of Pan-Kla were detected by Western blot ( n = 3 biological replicates). b Volcano plot depicting the differential gene expression between lactate-treated NB4 cells (Lac) and controls ( n = 3 biological replicates). Upregulated genes are shown in purple, and downregulated genes are shown in green (adjusted p -values < 0.05 and | log 2 (fold change) |>2). c Bubble chart showing gene ontology terms enriched in upregulated genes ( n = 3 biological replicates). d Heatmap showing the expression of differential genes in the coagulation pathway from RNA-seq data ( n = 3 biological replicates). e , f NB4 and MOLM13 cells were exposed to 20 mM lactate, followed by qPCR and Western blot to assess PAI-1 expression ( n = 3 biological replicates). g Cells were tested for plasmin generation with plasminogen and the substrate S2251, the absorbance (Abs) was continuously measured at 405 nm ( n = 3 biological replicates). h Fibrinolysis assays were conducted in lactate-treated NB4 and MOLM13 cells, fibrinolysis rate was recorded based on turbidity at 405 nm, dashed line represents 50% lysis of the clot ( n = 3 biological replicates). i TAT complex in plasma was measured using an ELISA kit ( n = 6 mice per group). j Representative immunofluorescence images showing Pan-Kla and PAI-1 expression in mouse bone marrow cells (scale bar=25 μm) ( n = 6 mice per group). k PAI-1 levels in mouse plasma measured using a commercial ELISA kit ( n = 6 mice per group). l Representative immunofluorescence images of PAI-1 content (yellow) in thrombi from thrombotic IVC segments of mice (scale bar = 200 μm) ( n = 6 mice per group). m ELISA quantification of t-PA/PAI-1 complex levels in mouse plasma ( n = 6 mice per group). Data were analyzed by two-tailed Wald test with Benjamini-Hochberg multiple comparisons ( b ), two-tailed hypergeometric test with Benjamini-Hochberg multiple comparisons ( c ), two-tailed Student’s t test ( e , i , l , m ), two-way ANOVA with Sidak’s multiple comparisons test ( g , h ), two-tailed Mann-Whitney test ( k ). Data are presented as mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Patients’ plasma PAI-1 levels were measured using a commercial ELISA kit (BOSTER, China).

Techniques: Western Blot, Gene Expression, Expressing, Coagulation, RNA Sequencing, Lysis, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Hexokinase 2-mediated histone H3K18la promotes PAI-1-dependent thrombosis in acute myeloid leukemia via tumor-endothelial crosstalk

doi: 10.1038/s41467-025-65259-0

Figure Lengend Snippet: a The outline depicts the steps of the animal experiments (by figdraw.com). b NGG mice were injected with sh HK2 or shNC AML cells via tail vein, and leukemia engraftment was confirmed by bioluminescence imaging ( n = 6 mice per group). c Plasma lactate was measured using a Lactate Assay Kit ( n = 6 mice per group). d Representative images and weights of thrombi isolated from mice 48 hours after IVC stenosis ( n = 6 mice per group). e Immunofluorescence staining of H3K18la and PAI-1 in mouse bone marrow cells (scale bar=25 μm) ( n = 6 mice per group). f Levels of PAI-1 in mice plasma were measured by ELISA ( n = 6 mice per group). g Representative immunofluorescence images of PAI-1 (yellow) expression in thrombi from mouse IVC (scale bar = 200 μm) ( n = 6 mice per group). h Plasma t-PA/PAI-1 complex levels were determined by ELISA ( n = 6 mice per group). shNC, negative control; sh HK2 , shRNA targeting HK2 . Data were analyzed by two-tailed Student’s t test ( b , c , g , h ), two-tailed Mann-Whitney test ( d , f ). Data are presented as mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Patients’ plasma PAI-1 levels were measured using a commercial ELISA kit (BOSTER, China).

Techniques: Injection, Imaging, Clinical Proteomics, Lactate Assay, Isolation, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Negative Control, shRNA, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Hexokinase 2-mediated histone H3K18la promotes PAI-1-dependent thrombosis in acute myeloid leukemia via tumor-endothelial crosstalk

doi: 10.1038/s41467-025-65259-0

Figure Lengend Snippet: a The outline depicts the steps of the animal experiments (by figdraw.com). b Bioluminescence images were acquired after leukemia implantation in mice before IVC stenosis. c Plasma lactate was measured using a Lactate Assay Kit. d Representative images and thrombus weights from mice 48 hours after IVC stenosis surgery. e Immunofluorescence staining of H3K18la and PAI-1 in mouse bone marrow cells (scale bar = 25 μm). f Immunofluorescence staining was performed to evaluate the expression of H3K18la and PAI-1 in the IVC tissues of thrombus-forming segment. (CD31, an endothelial cell marker; scale bar = 50 μm). g, h The levels of PAI-1 and t-PA/PAI-1 complex in mouse plasma were detected using ELISA kits. i Representative immunofluorescence images of PAI-1 (yellow) expression in thrombi from mouse IVC (scale bar = 200 μm). b-i n = 6 mice per group. Data were analyzed by one-way ANOVA with Dunnett’s multiple comparisons test. Data are presented as mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Patients’ plasma PAI-1 levels were measured using a commercial ELISA kit (BOSTER, China).

Techniques: Clinical Proteomics, Lactate Assay, Immunofluorescence, Staining, Expressing, Marker, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine

Article Title: Soluble urokinase plasminogen activator receptor promotes endoplasmic reticulum stress and apoptosis susceptibility through RAGE in sepsis acute kidney injury

doi: 10.1186/s10020-025-01352-w

Figure Lengend Snippet: The association between suPAR and acute kidney injury among ICU patients. A Comparison of serum suPAR levels at enrollment among patients in the three groups, ** P < 0.01. B After excluding the patients with pre-existing AKI at the time of enrollment, the ROC analysis was performed on the suPAR levels at enrollment, with the dichotomous variable being the occurrence of AKI. The area under the curve was 0.73. C - E . Correlations between the serum suPAR level at enrollment and the serum creatinine levels at the time of enrollment, 24 h after enrollment, and 48 h after enrollment, respectively

Article Snippet: The suPAR ELISA kit was provided by Elabscience (Cat No: E-EL-H2584c).

Techniques: Comparison

Journal: Molecular Medicine

Article Title: Soluble urokinase plasminogen activator receptor promotes endoplasmic reticulum stress and apoptosis susceptibility through RAGE in sepsis acute kidney injury

doi: 10.1186/s10020-025-01352-w

Figure Lengend Snippet: Increased suPAR/uPAR protein and mRNA levels in the kidneys of SA-AKI mice. A Serum levels of SCr, BUN, and suPAR in mice injected with saline (100 μL), LPS (10 mg/kg, 24 h), or subjected to CLP (24 h) ( n = 6 mice/group). B- C Representative images of PAS staining, TUNEL staining, and uPAR immunohistochemical staining in kidney tissues across experimental groups, along with quantitative analyses ( n = 3 mice/group; 3–10 fields/mouse). D - E uPAR immunofluorescence staining and quantitative assessment were performed in different groups of mice, with LTL staining used to visualize the brush border of renal tubular epithelial cells (White arrows: brush border discontinuity) ( n = 3 mice/group; 3 fields/mouse). F Protein expression levels of suPAR/uPAR in kidney tissues from different groups ( n = 3 mice/group; Mann–Whitney U test). G mRNA expression levels of uPAR in kidney tissues from different groups ( n = 6 mice/group). Data were presented as mean ± SEM. Statistical significance was determined by one-way ANOVA or unpaired two-tailed Student's t-test unless noted, with P < 0.05 considered significant (* P < 0.05, ** P < 0.01)

Article Snippet: The suPAR ELISA kit was provided by Elabscience (Cat No: E-EL-H2584c).

Techniques: Injection, Saline, Staining, TUNEL Assay, Immunohistochemical staining, Immunofluorescence, Expressing, MANN-WHITNEY, Two Tailed Test

Journal: Molecular Medicine

Article Title: Soluble urokinase plasminogen activator receptor promotes endoplasmic reticulum stress and apoptosis susceptibility through RAGE in sepsis acute kidney injury

doi: 10.1186/s10020-025-01352-w

Figure Lengend Snippet: suPAR promotes apoptosis susceptibility in renal tubular epithelial cells through ER stress and oxidative stress. A Protein levels of P-p65, p65, BAX, and BCL-2 in HK-2 cells treated with suPAR (10 ng/mL) at different time points. B Representative images of transmission electron microscope for endoplasmic reticulum in tubular epithelial cells (The black arrow refers to the endoplasmic reticulum). Scale bar: 0.5 μm. C Protein levels of P-PERK, PERK, P-eIF2α, ATF4, CHOP, Nrf2, and HO-1 in HK-2 cells treated with suPAR (10 ng/mL) at different time points. D mRNA expression levels of eIF2α in HK-2 cells at different time points. E Intracellular ROS levels in HK-2 cells at different time points. Data were presented as mean ± SEM from three independent experiments ( n = 3). Statistical significance was determined by Kruskal–Wallis test with Dunn's post hoc analysis, with P < 0.05 considered significant (* P < 0.05, ** P < 0.01)

Article Snippet: The suPAR ELISA kit was provided by Elabscience (Cat No: E-EL-H2584c).

Techniques: Transmission Assay, Microscopy, Expressing

Journal: Molecular Medicine

Article Title: Soluble urokinase plasminogen activator receptor promotes endoplasmic reticulum stress and apoptosis susceptibility through RAGE in sepsis acute kidney injury

doi: 10.1186/s10020-025-01352-w

Figure Lengend Snippet: suPAR mediates signal transduction through RAGE binding. A Protein levels of P-PERK, PERK, P-eIF2α, ATF4, CHOP, Nrf2, BAX and BCL-2 of HK-2 cells treated with suPAR(10 ng/ml) and/or FPS—ZM1(100 nM) or TLR4—IN—C34(10 μM). B The schematic diagram of the Protein docking between uPAR and RAGE. The ARG-116, SER-117, GLU-37, GLU-39, GLY-10, and ASP-11 of uPAR form hydrogen bonds with the GLN-24, ASN-81, GLY-10, THR-109, and LYS-107 of RAGE respectively. The lengths of the hydrogen bonds are 2.5 Å, 2.6 Å, 2.4 Å, 3.2 Å, 3.3 Å, and 3.5 Å respectively. C Direct suPAR-RAGE interaction confirmed by pull-down assay coupled with western blotting. D Representative confocal images showed the colocalization of suPAR (green) and RAGE (red) in HK-2 cells. Scale bar: 25 μm. Data were presented as mean ± SEM from three independent experiments ( n = 3). Statistical significance was determined by Kruskal–Wallis test with Dunn's post hoc analysis, with P < 0.05 considered significant (* P < 0.05, ** P < 0.01)

Article Snippet: The suPAR ELISA kit was provided by Elabscience (Cat No: E-EL-H2584c).

Techniques: Transduction, Binding Assay, Pull Down Assay, Western Blot