Journal: eBioMedicine

Article Title: Reprogramming offspring liver health: maternal indole supplementation as a preventive strategy against MASLD

doi: 10.1016/j.ebiom.2025.106098

Figure Lengend Snippet: Gut microbiota partially mediates the protective effects of perinatal Ind and I3A exposure on hepatic fibrosis and ceramide metabolism. ( a ) Body weight monitored weekly in FMT recipients. Data are mean ± SEM, n = 5/group. ( b ) Histological examination of liver sections from FMT recipients by H&E, picrosirius red (PSR), and immunohistochemistry of COL3A1 to quantify fibrosis. Representative images are shown from n = 5 mice/group. Magnification: 20×; scale bar: 100 μm. Quantification of PSR positive area ( c ) and COL3A1 ( d ). Data are mean ± SEM; n = 5/group. ∗p < 0.05 and ∗∗p < 0.01 vs. WD by one-way ANOVA with Dunnett's test. ( e ) Hepatic long-chain (LC) and very long-chain (VLC) ceramide levels by LC-MS. Data are mean ± SEM; n = 5/group.∗p < 0.05 vs. WD by one-way ANOVA with Dunnett's test. Hepatic gene expression of ceramide synthesis genes ( f ), ceramide degradation genes ( g ), and Ahr and its canonical targets ( h ) by qPCR, normalised to Rn18s . Data are mean ± SEM; n = 5/group. ∗p < 0.05 vs. WD by one-way ANOVA with Dunnett's test. ( i ) Gene expression of tight junction genes in ileum from FMT recipients by qPCR, normalised to Rn18s . Data are mean ± SEM; n = 5/group. ∗p < 0.05 vs. WD by Kruskal–Wallis with Dunn's test.

Article Snippet: Sections were incubated in 5% goat serum for 30 min, primary antibody COL1A1 (liver; 1:200; Cell Signalling Technologies [CST] Cat# 72026, RRID: AB_2904565 ), COL3A1 (liver; 1:1000; Proteintech Cat# 22734-1-AP, RRID: AB_2879158 ) or AHR (colon; 1:200; Novus Cat# NB100-2289, RRID: AB_10002581 ) for 60 min, followed by poly-HRP IgG secondary antibody.

Techniques: Immunohistochemistry, Liquid Chromatography with Mass Spectroscopy, Gene Expression

Journal: bioRxiv

Article Title: ADGRG1 blockade drives astrocyte positioning into the extracellular matrix to reduce scar size

doi: 10.64898/2025.12.18.695114

Figure Lengend Snippet: (A) UMAP of cell states (cell types) in the injured spinal cord snRNA-seq (n = 151712) datasets. Astro: Astrocytes, ABC: Arachnoid barrier cells, Endo: Endothelial cells, VLMC: vascular leptomeningeal cells. (B) Stacked bar plot showing the temporal changes in the relative proportions of astrocyte subclusters. dpi: days post-injury. (C) Dot plots showing the snRNA-seq analysis results concerning cell-cell interactions at 4 dpi. Color indicates co-expression of ligand and astrocyte receptor genes; transparency reflects the interaction score; presence of a red outline indicates statistical significance. P-values < 0.05 were considered significant. (D) Circos plots showing the number of significant and strong cell interactions via type III collagen in the injured spinal cord at 4 dpi. Numbers above the arrows indicate the number of pathways that were both significant and strong for each cell–cell pair. Arrows outlined in red indicate the COL3A1-ADGRG1 pathway. P-values < 0.05 were considered significant. Interaction scores > 70 were considered strong. (E) Line plots showing temporal changes in Type III collagen-ADGRG1 pathway interaction scores in the spinal cord. (F) 3D-spatial transcriptomics of the injured spinal cord at 7 dpi, highlighting type III collagen–secreting cells and neighboring astrocytes. The figure depicts the spatial localizations of Astro_1 (green), VLMC_1 (blue), and ABC_2 (orange).

Article Snippet: The sections were then stained with primary antibodies against GFAP (1:500; astrocyte marker, rat; 13-0300; Invitrogen), Type III collagen (1:500; rabbit; Protein Tech), or ADGRG1 (1:500; rabbit; bs-11848R; Bioss).

Techniques: Expressing

Journal: bioRxiv

Article Title: ADGRG1 blockade drives astrocyte positioning into the extracellular matrix to reduce scar size

doi: 10.64898/2025.12.18.695114

Figure Lengend Snippet: (A) Representative images showing GFAP (green) and type III collagen (red) staining of spinal cords at 14 dpi receiving PBS (control group) or DHM (DHM inj group). The white dotted line indicates the border of the glial scar. Scale bars: 500 μm and 50 μm (inset). (B) Three-dimensional images of the scar in the control group and in the DHM inj group (red: type III collagen-positive volume surrounded by GFAP-positive area). R indicates the rostral side, D indicates the dorsal side, and L indicates the left side. Grid size: 500 μm. (C) Quantitative analysis of the area of astrocytes inside type III collagen in the control group and in the DHM inj group (n = 6). (D) Quantitative analysis of scar volume between the control group and DHM inj group (n = 6). (E) Neuronal tracing images showing tdTomato (red)-positive axons and GFAP (green)-positive astrocytes in the control group and in the DHM inj group. Scale bars: 500 μm and 50 μm (inset). (F) Quantitative analysis of axons in the control group and in the DHM inj group (n = 8). (G-I) Changes in Il-6 , Il-1β , and Tnf-α gene expression between the control group and the DHM inj group at 7 dpi (n = 5). (J) The time course of the BMS score after SCI in the control group and in the DHM inj group (n = 10). (K) Representative images showing GFAP (red), type III collagen (cyan), and GFP (green, inset) staining of spinal cords at 14 dpi, treated with control AAV (control AAV group) or AAV- Gfap - shAdgrg1 ( Adgrg1 K.D group). Scale bars: 200 μm and 50 μm (inset). (L) Quantitative analysis of the area of astrocytes inside type III collagen in the control AAV group and in the Adgrg1 K.D group (n = 5). (M) Quantitative analysis of scar volume between the control AAV group and the Adgrg1 K.D group (n = 5). (N) The time course of the BMS score after SCI in the control AAV group and in the Adgrg1 K.D group (n = 10). *P < 0.05, **P < 0.005, unpaired t-test (C, D, F-I, and L-M), two-way ANOVA (J and N). Data represent the mean ± s.e.m (C-D, F-J, and L-N).

Article Snippet: The sections were then stained with primary antibodies against GFAP (1:500; astrocyte marker, rat; 13-0300; Invitrogen), Type III collagen (1:500; rabbit; Protein Tech), or ADGRG1 (1:500; rabbit; bs-11848R; Bioss).

Techniques: Staining, Control, Gene Expression

Journal: bioRxiv

Article Title: ADGRG1 blockade drives astrocyte positioning into the extracellular matrix to reduce scar size

doi: 10.64898/2025.12.18.695114

Figure Lengend Snippet: (A) In vitro scratch injury of monolayer-cultured astrocytes on poly-L-lysine coating, type III collagen coating, or type III collagen coating with DHM. Scale bar: 100 μm. (B) Quantitative analysis of the number of migrating cells among the three groups (n = 5 per group). (C) Representative images and migration plots of astrocytes from cell tracking assay among the three groups. Scale bar: 50 μm. (D) Quantitative analysis of migration distance among the three groups. (E) Quantitative analysis of RHOA-GTP binding levels among the three groups. (F) In vitro cell migration assay of monolayer-cultured astrocytes on type III, type I, or type IV collagen coating. Control: astrocytes cultured on type III collagen with the stopper kept until fixation. Representative images depicting GFAP (green) and Hoechst (blue). Scale bar: 500 μm. (G) Quantitative analysis of the number of migrating cells among the three groups. *P < 0.05, **P < 0.005, one-way ANOVA (B, D, E, G). Data represent the mean ± s.e.m (B, D, G). Data represent minimum to maximum (E).

Article Snippet: The sections were then stained with primary antibodies against GFAP (1:500; astrocyte marker, rat; 13-0300; Invitrogen), Type III collagen (1:500; rabbit; Protein Tech), or ADGRG1 (1:500; rabbit; bs-11848R; Bioss).

Techniques: In Vitro, Cell Culture, Migration, Cell Tracking Assay, Binding Assay, Cell Migration Assay, Control

Journal: bioRxiv

Article Title: ADGRG1 blockade drives astrocyte positioning into the extracellular matrix to reduce scar size

doi: 10.64898/2025.12.18.695114

Figure Lengend Snippet: (A) Quantitative analysis of astrocyte process length cultured on poly-L-lysine, type III collagen, or type III collagen with DHM. (B) Quantitative analysis of migration speed among the three groups.

Article Snippet: The sections were then stained with primary antibodies against GFAP (1:500; astrocyte marker, rat; 13-0300; Invitrogen), Type III collagen (1:500; rabbit; Protein Tech), or ADGRG1 (1:500; rabbit; bs-11848R; Bioss).

Techniques: Cell Culture, Migration

Journal: bioRxiv

Article Title: ADGRG1 blockade drives astrocyte positioning into the extracellular matrix to reduce scar size

doi: 10.64898/2025.12.18.695114

Figure Lengend Snippet: (A) Representative images of hiPSC-derived astrocytes showing GFAP (green), ADGRG1 (red), and Hoechst (blue) staining. Scale bar: 20 μm. (B) Quantitative analysis of ADGRG1-positive hiPSC-derived astrocytes. (C) In vitro scratch injury of monolayer-cultured hiPSC-derived astrocytes on Geltrex coating, type III collagen coating, or type III collagen coating with DHM. Scale bar: 500 μm. (D) Quantitative analysis of the number of migrating cells among the three groups (n = 6 per group). (E) Migration plots from the cell tracking assay of hiPSC-derived astrocytes (n = 5 per group). The tick interval is 100 μm. (F) Quantitative analysis of the migration distance among the three groups. *P < 0.05, **P < 0.005, unpaired t-test (B), one-way ANOVA (D, F). Data represent the mean ± s.e.m (B, D, F).

Article Snippet: The sections were then stained with primary antibodies against GFAP (1:500; astrocyte marker, rat; 13-0300; Invitrogen), Type III collagen (1:500; rabbit; Protein Tech), or ADGRG1 (1:500; rabbit; bs-11848R; Bioss).

Techniques: Derivative Assay, Staining, In Vitro, Cell Culture, Migration, Cell Tracking Assay