Journal: Reproductive Sciences

Article Title: Female Infertility and Risk for Later-Life Cardiovascular Disease: Lessons from a Mouse Model of Human Cardiovascular Disease

doi: 10.1007/s43032-025-02026-y

Figure Lengend Snippet: Immunohistochemical evaluation of uteri on day 4.5 post-coitus in SR-BI KO/ ApoeR61 h/h mice. Leptin receptor (Leptin R) ( a-c ), cyclooxygenase (COX) −2 ( d-f ), leukaemia inhibitory factor (LIF) ( g-i ) and phospho-signal transducer and activator of transcription (Stat) −3 ( j-l ) immunostaining with haematoxylin counterstaining in SR-BI KO/ ApoeR61 h/h mice with placebo ( b , e , h , k ) or probucol treatment ( c , f , i , l ). As a control, uteri from wild type mice ( a , d , g , j ) were evaluated. Scale bar = 100 µm

Article Snippet: The 5-μm paraffin-embedded tissue sections were immunostained using monoclonal rabbit anti-mouse phosphorylated signal transducer and activator of transcription-3 (p-Stat3) antibody (Tyr705) (#9145; Cell Signaling Technology, Tokyo, Japan) at 1:400 dilution, polyclonal rabbit anti-mouse leukaemia inhibitory factor (LIF) antibody (OABF00432; Aviva Systems biology, CA, US) at 1:400 dilution, polyclonal goat anti-mouse leptin receptor antibody (AF497; R&D Systems, MN, US) at 1:200 dilution, and polyclonal rabbit anti-mouse cyclooxygenase (COX) −2 antibody (#160,126; Cayman Chemical, MI, US) at 1:500 dilution according to the manufacturers’ instructions.

Techniques: Immunohistochemical staining, Immunostaining, Control

Journal: Experimental and molecular pathology

Article Title: Expression of free fatty acid receptor 2 in normal and neoplastic tissues.

doi: 10.1016/j.yexmp.2024.104902

Figure Lengend Snippet: Fig. 7. Double-labelling immunohistochemical analysis of free fatty acid receptor 2 (FFAR2) expression and the expression of gastric inhibitory polypeptide (GIP; A–C) in the human duodenum, glucagon-like peptide 1 (GLP-1) in the human jejunum (D–F), or peptide YY in the human duodenum (G–I). Labelling of FFAR2 was visualised using Cy3-conjugated goat anti-rabbit antibody (red). Labelling of GIP, GLP-1, or peptide YY was visualised using Alexa Fluor 488-conjugated donkey anti- mouse antibody. Overlapping expression is represented by orange/yellow colour. Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. Scale bar, 100 μm (A–I). Arrows in (A–C) indicate an enteroendocrine cell expressing both FFAR2 and GIP; arrowheads in (A–C) indicate an FFAR2-positive and GIP- negative enteroendocrine cell; arrows in (D–F) indicate GLP-1-positive and FFAR2-negative enteroendocrine cells; arrows in (G–I) indicate enteroendocrine cells expressing only FFAR2; and arrowheads in (G–I) indicate enteroendocrine cells expressing only peptide YY. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For double-labelling fluorescence immunohistochemistry, sections were incubated overnight at 4 ◦C with rabbit anti-FFAR2 0524 antibody (1:100 dilution) together with mouse monoclonal anti-insulin antibody (1:100 dilution; Abcam), mouse monoclonal anti-glucagon antibody (1:500 dilution; Sigma-Aldrich, St. Louis, MO, USA), rat monoclonal anti-somatostatin-14/28 antibody (1:300 dilution; Abcam, Cambridge, UK), guinea pig polyclonal anti-gastrin antibody (1:800 dilution; Origene, Rockville, MD, USA), rat monoclonal anti-ghrelin antibody (1:50; R&D Systems/biotechne, Wiesbaden, Germany), mouse monoclonal anti-serotonin antibody (1:50 dilution; DAKO, Glostrup, Denmark), mouse monoclonal anti-gastric inhibitory polypeptide (GIP) antibody (1:1500 dilution; Invitrogen, Waltham, MA, USA), mouse monoclonal anti-glucagon-like peptide-1 (GLP-1) antibody (1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA), or mouse monoclonal antipeptide YY antibody (1:100 dilution; Novus Biologicals/biotechne, Wiesbaden, Germany).

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: Cellular and Molecular Life Sciences

Article Title: Restraining of glycoprotein VI- and integrin α2β1-dependent thrombus formation by platelet PECAM1

doi: 10.1007/s00018-023-05058-2

Figure Lengend Snippet: Enhanced collagen-induced thrombus formation by PECAM1 blockage in the absence of integrin α2β1. Blood samples were preincubated with anti-α2β1 6F1 mAb (20 µg/mL) or vehicle, and then perfused during 3.5 min at 1000 s −1 over microspots of collagen-I ( A ) or collagen-III ( B ); microspots were post-coated with saline or inhibitory anti-PECAM1 mAb (clone WM59, mouse IgG1). Brightfield and multicolor microscopic images were taken at end stage with an EVOS-FL microscope and 60 × oil objective. Platelet labeling was withAF647 anti-CD62P mAb, FITC anti-fibrinogen mAb and AF658 annexin A5. Shown are representative images for per collagen surface with or without inhibitory anti-PECAM1 mAb ( n = 6–7). EVOS microscope; scale bars, 50 µm

Article Snippet: Control anti-PECAM-1.2 mAb (MABF-2034, clone MBC 78.2, mouse IgG) came from Sigma-Aldrich (Merck, Amsterdam, The Netherlands) Polyclonal anti-G6b-B Ab (PA5-23,300, rabbit IgG), inhibitory anti-FcγRIIA mAb (clone IV.3, mouse IgG2), and mouse IgG1 isotype control came from ThermoFisher Scientific (Eindhoven, The Netherlands).

Techniques: Saline, Microscopy, Labeling

Journal: Cellular and Molecular Life Sciences

Article Title: Restraining of glycoprotein VI- and integrin α2β1-dependent thrombus formation by platelet PECAM1

doi: 10.1007/s00018-023-05058-2

Figure Lengend Snippet: Selective effect of inhibitory anti-PECAM1 mAb in rescue of collagen-induced thrombus formation upon α2β1 inhibition. Blood samples were preincubated with 6F1 mAb (20 µg/mL) or vehicle, and then perfused during 3.5 min at 1000/s over microspots of collagen I, collagen III or collagen IV. Microspots were post-coated with saline, inhibitory anti-PECAM1 mAb (IgG1), mouse isotype control IgG1, or a control anti-PECAM mAb (clone MBC 78.2), as indicated. Multicolor microscopic images (EVOS-FL microscope, 60 × oil objective) were analyzed for eight parameters (Suppl. Table 1), which were univariate scaled. A , B Subtraction heatmaps for comparing per collagen type the effects of integrin α2β1 blockage (by 6F1 mAb) on parameters: P1, platelet deposition; P2, thrombus multilayer size; P3, thrombus morphological score; P4, thrombus multilayer score; P5, thrombus contraction score; P6, P-selectin expression; P7, integrin αIIbβ3 activation; P8, phosphatidylserine exposure. (A) Heatmapped effects of the 6F1 mAb on microspots of a collagen + inhibitory anti-PECAM1 mAb or the collagen + control IgG. B Heatmapped effects of 6F1 mAb on microspots of a collagen + control anti-PECAM1 mAb. Means ± SD ( n = 3–7), tested for significance with one-way ANOVA. For color coding, a filter was applied of p < 0.05. Raw data are given in Suppl. Datafile 1

Article Snippet: Control anti-PECAM-1.2 mAb (MABF-2034, clone MBC 78.2, mouse IgG) came from Sigma-Aldrich (Merck, Amsterdam, The Netherlands) Polyclonal anti-G6b-B Ab (PA5-23,300, rabbit IgG), inhibitory anti-FcγRIIA mAb (clone IV.3, mouse IgG2), and mouse IgG1 isotype control came from ThermoFisher Scientific (Eindhoven, The Netherlands).

Techniques: Inhibition, Saline, Microscopy, Expressing, Activation Assay