Journal: Pathophysiology

Article Title: Metabolic Syndrome-Driven Changes in Cardiac Lymphatic Endothelium: mRNA Expression and Emerging Questions

doi: 10.3390/pathophysiology33010004

Figure Lengend Snippet: LVs localized subepicardially and perivascularly in left ventricles of hearts from control ( a – h ) and db/db ( i – p ) mice. Most of the lymphatic vessels express all three markers: Lyve-1 (red), Pdpn (white) and CD31 (green) (merge; yellow arrows); few Pdpn-positive cells do not express Lyve-1 or CD31 (merge; white arrows). Merged panels ( d , h , l , p ) include DAPI staining of nuclei (blue). ( m – p ) shows zoomed region of ( i – l ). ( q ) shows relative quantification of mRNA level for Pdpn. ( r ) presents the percentage of Pdpn-positive LVs among all cardiac LVs, as shown in representative confocal microscope images. Scale bars—100 µm. Abbreviations: LV—lymphatic vessel, Pdpn—podoplanin. Statistically significant differences ( p -value < 0.05) are marked (*).

Article Snippet: Sections were stained with a cocktail of various combinations of antibodies to different antigens: Lyve-1 (R&D Systems, cat. no. MAB2125; Angiobio, San Diego, CA, USA, cat. no. 11-034, final concentration 1:300), Podoplanin (R&D Systems, cat. no. AF3244, final concentration 1:50), CD31 (BD Biosciences, cat. no. 550274, final concentration 1:100), CD31 (R&D Systems, cat. no. AF3628, final concentration 1:100), Collagen III (Novusbio, Centennial, CO, USA, cat. no. NBP1-26547, final concentration 1:10), Collagen I (Abcam, Cambridge, UK, cat. no. ab34710, final concentration 1:150), SMA (Abcam, cat. no. ab21027, final concentration 1:100), syndecan 4 (Invitrogen, Waltham, MA, USA, cat. no. PA1-32485, final concentration 1:40) diluted in PBS with 5% donkey serum for 1 h at room temperature or for 12 h in 4 °C.

Techniques: Control, Staining, Quantitative Proteomics, Microscopy

Journal: Pathophysiology

Article Title: Metabolic Syndrome-Driven Changes in Cardiac Lymphatic Endothelium: mRNA Expression and Emerging Questions

doi: 10.3390/pathophysiology33010004

Figure Lengend Snippet: Myocardial perivascular LVs surrounded by collagen I and collagen III in control ( a – h ) and db/db ( i – p ) mice. Merged panels ( d , h , l , p ) include DAPI staining of nuclei (blue). ( e – h ) shows zoomed region of ( a – d ). ( m – p ) shows zoomed region of ( i – l ). Scale bar—50 µm.

Article Snippet: Sections were stained with a cocktail of various combinations of antibodies to different antigens: Lyve-1 (R&D Systems, cat. no. MAB2125; Angiobio, San Diego, CA, USA, cat. no. 11-034, final concentration 1:300), Podoplanin (R&D Systems, cat. no. AF3244, final concentration 1:50), CD31 (BD Biosciences, cat. no. 550274, final concentration 1:100), CD31 (R&D Systems, cat. no. AF3628, final concentration 1:100), Collagen III (Novusbio, Centennial, CO, USA, cat. no. NBP1-26547, final concentration 1:10), Collagen I (Abcam, Cambridge, UK, cat. no. ab34710, final concentration 1:150), SMA (Abcam, cat. no. ab21027, final concentration 1:100), syndecan 4 (Invitrogen, Waltham, MA, USA, cat. no. PA1-32485, final concentration 1:40) diluted in PBS with 5% donkey serum for 1 h at room temperature or for 12 h in 4 °C.

Techniques: Control, Staining

Journal: STAR Protocols

Article Title: Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing

doi: 10.1016/j.xpro.2025.104296

Figure Lengend Snippet: 3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Article Snippet: Collagen III antibody, anti-human, PE, REAdye_lease , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-357 RRID: AB_2905171.

Techniques: Imaging, Comparison, Selection, Staining

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