sheep fap antibody  (R&D Systems)

Journal: bioRxiv

Article Title: Intercellular interaction between FAP fibroblasts and CD150 inflammatory monocytes mediates fibro-stenosis in Crohn’s disease

doi: 10.1101/2023.05.04.539383

Figure Lengend Snippet: ( A ) Experimental workflow showing FACS-sorting gating strategy and setup for in vitro ECM production. ( B ) Heatmap showing relative expression of FAP, TWIST1 and type III collagen in CCD-18Co fibroblast treated with supernatant from FACS sorted myeloid subsets (n= 5 individual patients). ( C ) Heatmap showing extracellular deposition of FAP, type I collagen and type III collagen in monocyte-stimulated CCD-18Co fibroblasts. ( D ) Heatmap showing relative protein expression of FAP, TWIST1 and type II collagen in primary ileal fibroblasts after being stimulated by selected cytokine combinations, predicted by NicheNet. ( E ) Experimental workflow. ( F ) Immunofluorescence staining (10× image) and ( G ) Bar plot showing relative expression of fibronectin and type IV collagen in pro-fibrotic cues -stimulated CCD-18Co fibroblasts after TWIST1 inhibition. Data are shown as bar plots with SEM. Statistically significant differences were determined using a one-way ANOVA test corrected with Tukey’s multiple comparisons test (* p <0.05, ** p <0.01, *** p <0.005, **** p <0.001).

Article Snippet: Then the cells were fixed with ice-cold acetone (Chem-Lab) and labelled with sheep anti-human FAP antibody (1:40, AF3715, R&D Systems), mouse anti-human TWIST1 antibody (1:100, clone 2F8E7, Invitrogen), goat anti-human collagen III alpha 1 antibody (1:500, NBP1-26547, Novus Biologicals), ATTO 490LS-conjugated phalloidin (1:200, 14479, Sigma-Aldrich), Alexa Fluor 488-conjugated donkey anti-sheep IgG antibody (A-11015, Invitrogen), Cy3-conjugated donkey anti-goat IgG antibody (705-165-003, Jackson ImmunoResearch) and Cy5-conjugated donkey anti-mouse IgG antibody (715-175-151, Jackson ImmunoResearch).

Techniques: In Vitro, Expressing, Immunofluorescence, Staining, Inhibition

Journal: bioRxiv

Article Title: Intercellular interaction between FAP fibroblasts and CD150 inflammatory monocytes mediates fibro-stenosis in Crohn’s disease

doi: 10.1101/2023.05.04.539383

Figure Lengend Snippet: ( A ) Heatmap showing relative expression of NicheNet-predicted ligands expressed by FACS-sorted myeloid cell subsets (n=4). ( B ) Immunofluorescence staining (25× image) and ( C ) Heatmap showing relative expression of FAP, TWIST1 and type III collagen in monocyte-stimulated CCD-18Co fibroblasts. ( D ) Immunofluorescence staining (10× image) and ( E ) Bar plot showing relative expression of FAP, type I and type III collagen in monocyte-stimulated CCD-18Co fibroblasts. Data are shown as bar plots with SEM. Statistically significant differences were determined using a one-way ANOVA test corrected with Tukey’s multiple comparisons test (*** p <0.005, **** p <0.001). ( F ) Immunofluorescence staining (25× image) and ( G ) Heatmap showing relative expression of FAP, TWIST1 and type III collagen in pro-fibrotic cues -stimulated CCD-18Co fibroblasts. ( H ) Immunofluorescence staining (10× image) and ( I ) Bar plot showing relative expression of FAP, type I and type III collagen in pro-fibrotic cues -stimulated CCD-18Co fibroblasts after TWIST1 inhibition. Data are shown as bar plot with SEM. ( J ) Immunofluorescence staining (10x image) and ( K ) quantitative analysis (mean fluorescence intensity) of FAP and PDPN pro-fibrotic cues -stimulated iPSC-derived intestinal organoids with or without Harmine. Data are shown as bar plot with SEM. Statistically significant differences were determined using a one-way ANOVA test corrected with Tukey’s multiple comparisons test (* p <0.05, ** p <0.01, *** p <0.005, **** p <0.001).

Article Snippet: Then the cells were fixed with ice-cold acetone (Chem-Lab) and labelled with sheep anti-human FAP antibody (1:40, AF3715, R&D Systems), mouse anti-human TWIST1 antibody (1:100, clone 2F8E7, Invitrogen), goat anti-human collagen III alpha 1 antibody (1:500, NBP1-26547, Novus Biologicals), ATTO 490LS-conjugated phalloidin (1:200, 14479, Sigma-Aldrich), Alexa Fluor 488-conjugated donkey anti-sheep IgG antibody (A-11015, Invitrogen), Cy3-conjugated donkey anti-goat IgG antibody (705-165-003, Jackson ImmunoResearch) and Cy5-conjugated donkey anti-mouse IgG antibody (715-175-151, Jackson ImmunoResearch).

Techniques: Expressing, Immunofluorescence, Staining, Inhibition, Fluorescence, Derivative Assay

Journal: bioRxiv

Article Title: Intercellular interaction between FAP fibroblasts and CD150 inflammatory monocytes mediates fibro-stenosis in Crohn’s disease

doi: 10.1101/2023.05.04.539383

Figure Lengend Snippet: ( A ) Picrosirius red staining and ( B) Bar plot showing collagen deposition in mouse colon of chronic DSS colitis after Harmine treatment. Data are shown as bar plots with SEM. Statistically significant differences were determined using a one-way ANOVA test corrected with Tukey’s multiple comparisons test (* p <0.05, ** p <0.01). ( C ) Immunofluorescence staining for FAP and TWIST1 expression in mouse colon of chronic DSS colitis after Harmine treatment. Original image composed of stitched 25× images.

Article Snippet: Then the cells were fixed with ice-cold acetone (Chem-Lab) and labelled with sheep anti-human FAP antibody (1:40, AF3715, R&D Systems), mouse anti-human TWIST1 antibody (1:100, clone 2F8E7, Invitrogen), goat anti-human collagen III alpha 1 antibody (1:500, NBP1-26547, Novus Biologicals), ATTO 490LS-conjugated phalloidin (1:200, 14479, Sigma-Aldrich), Alexa Fluor 488-conjugated donkey anti-sheep IgG antibody (A-11015, Invitrogen), Cy3-conjugated donkey anti-goat IgG antibody (705-165-003, Jackson ImmunoResearch) and Cy5-conjugated donkey anti-mouse IgG antibody (715-175-151, Jackson ImmunoResearch).

Techniques: Staining, Immunofluorescence, Expressing