db1976 gc39628  (GlpBio Technology Inc)

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: Characterization and Targeting Ability of D-NVs and D-FNVs. ( A ) Representative immunofluorescence images showing FOLR2 and CD68 expression in the aortic root sections from mice fed with a chow diet or a high-fat diet. ( B ) TEM image of D-FNVs, confirming their spherical morphology. Scale bar: 100 nm. ( C ) Western blot analysis of scavenger receptor A (SR-A) and Toll-like receptor 4 (TLR4) expression in macrophage-derived nanovesicles (D-NVs and D-FNVs). ( D - E ) Particle size and zeta potential of D-NVs and D-FNVs. ( F ) Encapsulation efficiency of D-NVs and D-FNVs. ( G ) Loading efficiency of D-NVs and D-FNVs. ( H ) Stability of D-NVs and D-FNVs evaluated by monitoring particle size and zeta potential over 10 days. ( I ) Release profile of DB1976 from D-FNVs under different pH conditions (pH 7.4 and pH 6.5) at 37 °C. ( J ) Representative confocal fluorescence images of macrophages incubated with DiI-labeled D-NVs or D-FNVs (red) and stained with DiO (green). Scale bar: 50 μm. ( K ) Confocal images of macrophages treated with ox-LDL and subsequently incubated with DiI-labeled D-NVs or D-FNVs. Scale bar: 50 μm. ( L ) Confocal images of ox-LDL–treated macrophages incubated with DiI-labeled D-FNVs in the presence or absence of free folate. Scale bar: 50 μm. ( M – O ) Quantification of relative fluorescence intensity (RFI) per cell showing uptake of D-NVs and D-FNVs ( M ), uptake of D-NVs and D-FNVs in ox-LDL–treated macrophages ( N ), and competition assay with free folate ( O ). One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Immunofluorescence, Expressing, Western Blot, Derivative Assay, Zeta Potential Analyzer, Encapsulation, Fluorescence, Incubation, Labeling, Staining, Competitive Binding Assay, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: PU.1 is upregulated in macrophages of advanced atherosclerotic lesions and promotes inflammation through IL-1β/NF-κB signaling. ( A ) Boxplot showing expression levels of PU.1 in early versus advanced atherosclerotic plaques based on the GSE43292 dataset. ( B ) UMAP plot of single-cell RNA sequencing data depicting major immune and stromal cell populations in atherosclerotic lesions. ( C ) UMAP feature plot showing SPI1 (encoding PU.1) expression predominantly enriched in macrophages. ( D ) Representative immunofluorescence staining of PU.1 (green) and CD68 (red) in aortic root sections from chow diet– and high fat diet–fed mice. Nuclei were counterstained with DAPI (blue). Scale bar: 200 μm. ( E ) CUT&Tag analysis showing genome-wide binding of PU.1 in macrophages. Heatmap indicates PU.1 enrichment near transcription start sites (TSS). ( F ) Genomic distribution of PU.1 binding peaks identified by CUT&Tag. ( G – H ) Representative CUT&Tag tracks showing PU.1 binding at the promoters of pro-inflammatory cytokines. ( I ) Dual-luciferase reporter assay confirming the transcriptional activation of the IL-1β promoter by PU.1 overexpression (OE). ( J ) Western blot showing that PU.1 knockdown suppressed ox-LDL–induced IL-1β expression and NF-κB pathway activation (p-IκB and p-p65). ( K ) Western blot demonstrating that IL-1β knockdown reversed PU.1-induced NF-κB activation. ( L ) Western blot analysis showing that the PU.1 inhibitor DB1976 attenuated ox-LDL–induced IL-1β expression and NF-κB activation. ( M ) qRT-PCR analysis of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, MCP-1) in macrophages treated with ox-LDL with or without DB1976. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Expressing, RNA Sequencing, Immunofluorescence, Staining, Genome Wide, Binding Assay, Luciferase, Reporter Assay, Activation Assay, Over Expression, Western Blot, Knockdown, Quantitative RT-PCR, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: The anti-atherosclerotic effects of the D-FNVs. ( A - D ) BMDMs were co-treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs). The levels of IL-1β, IL-6, TNF-α, and MCP-1 in the supernatant were measured by ELISA. ( E , F ) Flow cytometry analysis and quantification of intracellular ROS levels in BMDM cells treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs), respectively, at 2 mM DB1976 for 24 h. ( G , H ) Flow cytometry analysis and quantification of apoptosis rates in BMDMs treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs) at 2 mM DB1976 for 24 h. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: Biodistribution and pharmacokinetics of D-NVs and D-FNVs in vivo. ( A ) Fluorescence imaging of DiD fluorescent signals in aortic tissues. ApoE −/− mice fed a high-fat diet for 3 months were intravenously administered DiD-labeled D-NVs or D-FNVs. Ex vivo fluorescence imaging of aortic tissues was performed using IVIS. ( B ) Quantitative analysis of DiD fluorescent signals in aortic tissues. ( C ) Confocal laser scanning microscopy (CLSM) images of DiD-labeled D-NVs and D-FNVs accumulated in atherosclerotic plaques of aortic root sections. Plaques are outlined by white dashed lines. Scale bar: 200 μm. ( D ) Biodistribution of D-NVs and D-FNVs in major organs (heart, liver, spleen, lung, and kidney) evaluated at 24 and 48 h after intravenous administration of DiD-labeled nanovesicles.( E ) DB1976 concentration in aortic tissues after administration of free DB1976, D-NVs, or D-FNVs. ( F ) Plasma pharmacokinetics of DB1976 following intravenous administration of free DB1976, D-NVs, or D-FNVs. ( G ) DB1976 distribution in major organs at 24 h after injection. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Drug discovery, In Vivo, Fluorescence, Imaging, Labeling, Ex Vivo, Confocal Laser Scanning Microscopy, Concentration Assay, Clinical Proteomics, Injection, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: Therapeutic efficacy of free DB976, D-NVs, D-FNVs on atherosclerosis ( A ) Schematic illustration of the development of an atherosclerotic mouse model and the treatment protocol with various formulations (Saline, free DB1976, D-NVs, and D-FNVs) at a dose of 2 mg/kg DB1976 per injection. ( B ) Representative photographs of En face Oil Red O (ORO)-stained aortas collected from atherosclerotic mice after treatment with the indicated formulations. ( C ) Quantitative analysis of the aortic lesion area, expressed as the mean ± SD. ( D - E ) Representative microphotographs and quantitative analysis of Oil Red O-stained aortic root sections from mice treated with different formulations. Scale bar: 500 μm. ( F ) Representative hematoxylin and eosin (H&E)-stained aortic root sections from the different treatment groups. Scale bar: 500 μm. ( G ) Quantification of the percentage of necrotic core area in aortic root plaques (mean ± SD). One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Drug discovery, Saline, Injection, Staining, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: D-FNVs ameliorated inflammation in an atherosclerotic mouse model. ( A , B ) Representative images of plaques within the aortic root subjected to immunofluorescent staining for the macrophage marker CD68. Scale bar: 100 μm. ( C – F ) Levels of IL-1β, IL-6, TNF-α, and MCP-1 in aortic tissues collected from atherosclerotic mice treated with various formulations (saline, DB1976, D-NVs, D-FNVs). ( G – J ) Levels of IL-1β, IL-6, TNF-α, and MCP-1 in blood serum collected from the same groups of atherosclerotic mice. The n values are all biological replicates. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, and *** p < 0.001. **** p < 0.0001

Article Snippet: Folic acid (HY-16637) and DB1976 (HY-135797 A) was acquired from MedChemExpress (China).

Techniques: Staining, Marker, Saline, Comparison, Standard Deviation

Journal: Nature Communications

Article Title: TLR4 + group 2 innate lymphoid cells contribute to persistent type 2 immunity in airway diseases

doi: 10.1038/s41467-025-62532-0

Figure Lengend Snippet: A Schematic diagram of the experimental study design for scATAC-seq of lung tissues from wide-type HDM sensitized mice. B Uniform manifold approximation and projection (UMAP) plots of sorted viable CD45 + Lin ⁻ ICOS + ST2 + cells from HDM-sensitized mice at day 45 and day 47. Each dot represents a single cell, with different ILC2 subsets shown in different colors. C Alteration of chromatin accessibility in the ILC2_c02 subset at day 47 versus day 45. The Y axis presents the log2 fold change of gene activity at day 47 versus day 45 and in library 2, while the X axis shows the log2 fold change in library 1. D Transcription factor binding motifs significantly enriched in the ILC2_c02 subset at day 47 versus day 45. P- values were calculated by a right-tailed hypergeometric test. E DNA accessibility tracing of the Tlr4 and Il1rl1 of different ILC2 subsets. Shaded boxes delineate increased chromatin accessibility. F Predicted motifs within Tlr4 promoter. G Motif footprinting trace showing binding activities of transcription factor JunB, Fos, PU.1 and MafK.

Article Snippet: To confirm the role of transcriptional factor in induction of TLR4 on ILC2s, the ILC2s were pre-treated with the transcription factor c-Fos / AP-1 inhibitor T-5224 (20 μM, MCE) and the PU.1 inhibitor DB1976 (100 nM, MCE) for 1 h prior to stimulation.

Techniques: Activity Assay, Binding Assay, Footprinting

Journal: Signal Transduction and Targeted Therapy

Article Title: Spatiotemporal transcriptomics elucidates the pathogenesis of fulminant viral myocarditis

doi: 10.1038/s41392-025-02143-9

Figure Lengend Snippet: Cd8 + effector T cells induced the damage of cardiomyocytes. a Changes in average predicted proportion of cardiomyocytes across the infected ventricle in the late stage. b The expression of apoptosis, necroptosis, and pyroptosis genes during disease progression in the snRNA-seq data. c UMAP embedding of cardiomyocytes of the snRNA-seq data colored by manually annotated clusters. d Cell proportion changes of cardiomyocytes at different time points. e Box plots of death score and heart contraction score among different cardiomyocyte subclusters. Scores were calculated by AddModuleScore function of the Seurat package. f The spatial colocation of contraction genes and T cell cytotoxicity genes. Green means the expression of cardiac contraction genes, red means the expression of cardiac T cell cytotoxicity genes, and yellow means the co-expression status. Scale bar: 500 μm. g Box plots of apoptosis, pyroptosis, and necroptosis scores between cardiomyocytes that are near to Cd8 + effector T cells and those distant to Cd8 + effector T cells. h The spatial colocation of virus - -death + cells and Cd8 + effector T cells. Green means the abundance of virus - -death + CMs, red means the abundance of Cd8 + effector T cells, and yellow means the colacation of the two cell types. Scale bar: 500 μm. i Expression trend of Ifng in T cells along different infection times (fitted with loess). The line shadow showing the 95% confidence interval. j Western blot images of representative death genes expression in the vehicle and IFN-γ treated mouse primary cardiomyocytes. k The top regulons that activated in virus - -death high CMs. l The expression of key transcriptional factors in the heart of patients with FM (n = 3 in HC group, n = 4 in FM group, data are represented as mean ± SEM). m The expression of Spi1 in the heart of FM mice and control (n = 5 per group, data are represented as mean ± SEM). n The expression of Spi1 in IFN-γ treated mouse primary cardiomyocytes (n = 4 per group, data are represented as mean ± SEM). o Correlation between response to interferon gamma score and Spi1 AUCell score of cardiomyocytes . p The mRNA level changes of death genes in IFN-γ and si-Spi1 treated mouse primary cardiomyocytes (n = 4 per group, data are represented as mean ± SEM) . q Protein level changes of death genes in IFN-γ and si-Spi1 treated mouse primary cardiomyocytes. Survival rate ( r ), H&E staining images ( s ), representative echocardiography images ( t ) and cardiac functions (ejection fraction and fraction shortening) ( u ) changes of FM and Spi1 inhibitor-treated mice. ( n = 5 per group, data are represented as mean ± SEM). Scale bar: 500 μm. Ven ventricle, BZ border zone, MR myocarditic region, HC healthy control, FM fulminant myocarditis

Article Snippet: Spi1 inhibitor DB1976 dihydrochloride (Cat. HY-135797A) and Anti-Mouse IFN gamma Antibody (H22) (Cat. HY-P99136) were purchased from MedChemExpress (New Jersey, USA) and were given after CVB3 injection.

Techniques: Infection, Expressing, Biomarker Discovery, Virus, Western Blot, Control, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: Spatiotemporal transcriptomics elucidates the pathogenesis of fulminant viral myocarditis

doi: 10.1038/s41392-025-02143-9

Figure Lengend Snippet: The treatment effects of IVIG on FM mice. a Schematic diagram of the workflow for drug administration. b PCA analysis of control, infection and treatment samples from 1 to 7dpi. c The level of inflammation score in control, infection and treatment samples from 1 to 7dpi. d The level of CVB3 in control, infection and treatment samples from 1 to 7dpi. e Heatmap showing the expression level of PC1 genes. f Violin plot shows the expression of PC1 genes in different spatial areas. g The proportion of Mono_Ccr2, Mac_IFNIC and Inflammatory_Mac of infection and treatment groups. h Differential strength of cellular interactions among cardiac macrophages and T cells in infection and treatment groups. The blue line indicates a decrease in the treatment group. i The expression of chemokines ( Ccl2 and Ccl4 ) by Inflammatory_Mac in infection and treatment groups at different stages. j The level of CVB3, the expression of Ccl2 and Ccr2 , and the distribution of Inflammatory_Mac and Cd8 + effector T cells on cardiac sections in treatment group in the late stage. k The proportion of Cd8 + effector T cells in infection and treatment groups. l The expression of cytotoxic molecules ( Fasl and Gzmb ) by Cd8 + effector T cells in infection and treatment groups at different stages. m Expression trend of Ifng in T cells of infection and treatment groups along different infection time (fitted with loess). The line shadow showing the 95% confidence interval. n The AUCell score of Spi1 in control, infection and treatment groups. PC principle component

Article Snippet: Spi1 inhibitor DB1976 dihydrochloride (Cat. HY-135797A) and Anti-Mouse IFN gamma Antibody (H22) (Cat. HY-P99136) were purchased from MedChemExpress (New Jersey, USA) and were given after CVB3 injection.

Techniques: Control, Infection, Expressing

Journal: Frontiers in Neuroscience

Article Title: Targeting transcription factor pu.1 for improving neurologic outcomes after spinal cord injury

doi: 10.3389/fnins.2024.1418615

Figure Lengend Snippet: DB1976 treatment reduces lipid droplet accumulation and cholesterol crystal formation in the injured spinal cord. (A) BODIPY staining of lipid droplets in the sagittal sections at 28 dpi in the control and DB1976 groups. (B) Polarized light detection of cholesterol crystals in the sagittal sections at 28 dpi in the control and DB1976 groups. (C,D) Quantitative analysis of the percentage of BODIPY + (A) or Crystal + (B) in the injured area of the control and DB1976 groups. n = 3 mice per group for immunofluorescence analysis and polarized light detection. **** p < 0.0001.

Article Snippet: DB1976 (GC39628, Glpbio, United States) was diluted with phosphate buffered saline (PBS) to a concentration of 1 μg/μl.

Techniques: Staining, Control, Immunofluorescence

Journal: Frontiers in Neuroscience

Article Title: Targeting transcription factor pu.1 for improving neurologic outcomes after spinal cord injury

doi: 10.3389/fnins.2024.1418615

Figure Lengend Snippet: DB1976 treatment reduces chronic inflammatory response and promotes the repair of tight junctions between endothelial cells in the injured area after SCI. (A) Immunofluorescence staining of CD68 (red) and PDGFRβ (green) in the sagittal sections at 28 dpi in the control and DB1976 groups. (B,C) Immunofluorescence staining of CD31 (red; blood vessels) and (B) CLN-5 (green; tight junction protein) or (C) ZO-1 (green; tight junction protein) in the sagittal sections at 28 dpi in the control and DB1976 groups. ROI represents the boxed region on the left. Arrowheads indicate CLN-5 + (B) or ZO-1 + (C) vessels, while arrows indicate CLN-5 – (B) or ZO-1 – (C) vessels. (D) Quantitative analysis of the percentage of CD68 + area in the sagittal sections at 28 dpi in the control and DB1976 groups. (E,F) Quantitative analysis of the percentage of CLN-5 + (B) or ZO-1 + (C) vessels in the lesion epicenter of the control and DB1976 groups at 28 dpi. n = 3 mice per group for immunofluorescence analysis. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: DB1976 (GC39628, Glpbio, United States) was diluted with phosphate buffered saline (PBS) to a concentration of 1 μg/μl.

Techniques: Immunofluorescence, Staining, Control

Journal: Frontiers in Neuroscience

Article Title: Targeting transcription factor pu.1 for improving neurologic outcomes after spinal cord injury

doi: 10.3389/fnins.2024.1418615

Figure Lengend Snippet: DB1976 treatment reduces fibrotic scar formation and promotes glial connectivity in the injured area after SCI. (A) Immunofluorescence staining of fibronectin (red) and PDGFRβ (green) in the sagittal sections at 28 dpi in the control and DB1976 groups. (B) Immunofluorescence staining of Collagen I (red) and PDGFRβ (green) in the sagittal sections at 28 dpi in the control and DB1976 groups. (C–E) Quantitative analysis of the percentage of PDGFRβ + , Fibronectin + , and Collagen I + areas at the injury area in the control and DB1976 groups. (F) Immunofluorescence staining of GFAP (green) and PDGFRβ (red) in the sagittal sections at 28 dpi in the control and DB1976 groups. ROI represents the boxed region on the left. (G) Quantitative analysis of the area of the lesion epicenter (GFAP – ) in the control and DB1976 groups. n = 3 mice per group for immunofluorescence analysis. ** p < 0.01, **** p < 0.0001.

Article Snippet: DB1976 (GC39628, Glpbio, United States) was diluted with phosphate buffered saline (PBS) to a concentration of 1 μg/μl.

Techniques: Immunofluorescence, Staining, Control