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, 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: 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