Journal: Science Advances

Article Title: A beneficial environment promotes immune resilience through epigenetic regulation

doi: 10.1126/sciadv.ady7317

Figure Lengend Snippet: ( A ) Airway resistance measured after increasing concentrations of nebulized methacholine to anesthetized mice. ( B ) Representative BAL differential counts and PAS staining images and ( C ) quantification of BAL eosinophilia and ( D ) volume of mucus within airway epithelium. ( E ) Reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis from whole-lung homogenates detecting the expression of Il13 gene. ( F ) Serum titers of anti-OVA–specific IgE. ( G ) Annotated immune cells were subsetted from whole-lung scRNA-seq and 2D plotted using uniform manifold approximation and projection for dimension reduction (UMAP). ( H ) OVA and OVA + FD immune cell UMAP representation overlaid. ( I ) DEG number comparing OVA versus OVA + FD groups. ( J ) Representative flow cytometry plots highlighting MDM and DC identification and ( K and L ) quantification of frequency of these cell types within parent gates. Results are plotted as means ± SEM, and difference between means were compared using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test ( n = 6 to 9).

Article Snippet: Mouse CCL8 (BioLegend, no. 446904), human CCL8 (BioLegend, no. 442204), OVA-specific IgE (Chondrex, no. 3004), and OVA-specific IgG1 (Chondrex, no. 3013) were used according to the supplier’s protocols.

Techniques: Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Flow Cytometry

Journal: Science Advances

Article Title: A beneficial environment promotes immune resilience through epigenetic regulation

doi: 10.1126/sciadv.ady7317

Figure Lengend Snippet: ( A ) Volcano plot showing DEGs in OVA versus OVA + FD MDMs, analyzed by model-based analysis of single-cell transcriptomics (MAST). ( B ) Ccl8 gene expression in OVA versus OVA + FD. ( C ) Mouse serum CCL8 measured by enzyme-linked immunosorbent assay. ( D ) Serum CCL8 levels in human patients with asthma versus healthy controls. ( E ) WT mice were sensitized to OVA and treated daily with anti-CCL8 antibody or isotype 6 hours before OVA challenge. Representative BAL Giemsa cytospins and PAS-stained lung sections are shown. ( F ) Quantification of BAL eosinophils and ( G ) volume of mucus. ( H ) Lung inflammatory eosinophils (CD45 + , Ly6g neg , CD125 int , SiglecF +, CD11c + , and CD101 + ) quantified by flow cytometry. ( I and J ) CCL8 gene expression in murine BMDMs and human monocytes after 48 hours of FD exposure followed by 6 hours of LPS stimulation. ( K ) Average gene expression from MHCII-related genes ( H2-Eb1 , H2-Ab1 , H2-Aa , and Ciita ) were down-regulated in OVA + FD MDMs. ( L ) MHCII mean fluorescence intensity (MFI) on lung MDMs (CD45 + , F4/80 + , Ly6G neg , CD11b + , and CCR2 + ). ( M ) HLA-DR MFI in human monocyte–derived APCs exposed to FD for 48 hours and LPS for 6 hours. ( N ) BMDMs pulsed with OVA 323–339 peptide after FD and LPS stimulation and then cocultured with naive CD4 + T cells. T cell proliferation assessed by CellTrace™ Violet (CTV) MFI-based division index. ( O ) Proliferation-associated gene ( Cenpe , Top2a , and Mki67 ) average expression within CD4 + T cell subset in OVA versus OVA + FD. Modules were clustered by KEGG/Reactome terms; antigen presentation node shows altered interaction strength. Results are plotted as means ± SEM, and difference between means were compared using one-way ANOVA followed by Tukey’s post hoc test ( n = 4 to 19).

Article Snippet: Mouse CCL8 (BioLegend, no. 446904), human CCL8 (BioLegend, no. 442204), OVA-specific IgE (Chondrex, no. 3004), and OVA-specific IgG1 (Chondrex, no. 3013) were used according to the supplier’s protocols.

Techniques: Single-cell Transcriptomics, Gene Expression, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry, Fluorescence, Derivative Assay, Expressing, Immunopeptidomics

Journal: Bioactive Materials

Article Title: Three-dimensional bioprinted hiHeps hepatorganoids with enhanced hepatic functions for the treatment of liver failure and promotion of liver regeneration

doi: 10.1016/j.bioactmat.2025.12.024

Figure Lengend Snippet: Zonation characterization of 3D bioprinted human hepatocyte organoids. A. Representative live/dead assay of hiHep cells in 2D and 3D culture. Scale bar: 200 μm. B. Quantification of cell viability of hiHep cells in 2D and 3D culture (n = 3 per group). C. Cell viability of 2D cultured hiHeps and 3DP-HHO assessed by CCK-8 assay (n = 3 per group). D. Morphological comparison of 2D cultured hiHeps, 3D bioprinted hiHeps organoid, and 3DP-HHO after 1 week/month of culture (Red arrow: liver cord-like structures formed via self-assembly, white arrow: hepatocyte spheroids spontaneously formed by hiHeps). Scale bars: 50 μm. E. Volcano plot of differentially expressed genes in 3DP-HHO compared to 2D cultured hiHeps; key hepatic genes are highlighted. F. KEGG enrichment analysis of up-regulated genes showing biological processes associated with hepatic function. G. Heatmap of zonation marker gene expression (pericentral vs. periportal hepatocyte landmarks) across HepRG, hiHeps, 3DP-HHO, and primary human hepatocytes (PHH). H. Schematic diagram of zonation map illustrating the spatial gene expression of pericentral (red) and periportal (green) markers in healthy liver. I. Representative immunofluorescence images and quantification of CYP3A4 (red), HAL (green), and DAPI (blue) in healthy mouse liver, 2D-cultured hiHeps and 3DP-HHO, demonstrating spatially organized metabolic domains. (White arrow: CYP3A4-positive zones, indicating pericentral hepatocytes, white pentagram: HAL-positive zones, indicating periportal hepatocytes). Scale bars: mouse liver, 100 μm; 3DP-HHO, 25 μm.

Article Snippet: CYP3A4 Mouse mAb, 1:500, 67110-1-Ig, Proteintech, China.

Techniques: Live Dead Assay, Cell Culture, CCK-8 Assay, Comparison, Marker, Gene Expression, Immunofluorescence