Journal: Communications Biology

Article Title: Neutrophil extracellular trap gene expression signatures identify prognostic and targetable signaling axes for inhibiting pancreatic tumour metastasis

doi: 10.1038/s42003-025-08440-x

Figure Lengend Snippet: A Images of representative human PDAC tumour tissue sections stained by immunofluorescence (IF) for citrullinated histone H3 (cit-H3) and myeloperoxidase (MPO) to identify neutrophils and neutrophil extracellular traps (NETs; arrows). Boxes, regions of interest (ROI) shown at higher magnification to the right. Scale bars: lower magnification, 50 μm; higher magnification, 10 μm. Images of representative human PDAC tumour tissue sections stained by IF showing co-localisation of B integrin-beta 1 (ITGB1) and integrin-linked kinase (ILK) and C coiled-coil domain-containing protein 25 (CCDC25) and ILK (arrows). Boxes, ROIs shown at higher magnification to the right. Scale bars: lower magnification, 50 μm ( B ), 100 μm ( C ); higher magnification, 20 μm (both panels). D Images of representative tissue sections from MIA PaCa-2 PDAC xenograft primary tumours stained by IF for MPO and cit-H3 to identify NETs (arrows). Scale bar, 20 μm. E Images of representative primary tumour tissue sections from the KPCY genetically engineered mouse model (GEMM) of PDAC stained by IF for MPO and cit-H3 to identify NETs (arrows). Scale bar, 10 μm. F Images of representative tissue sections of liver metastases from MIA PaCa-2 PDAC xenografts stained by IF for MPO and cit-H3 to identify NETs (arrows). Scale bar, 20 μm. G Images of representative tissue sections of liver metastases from the KPCY GEMM stained by IF for MPO to identify neutrophils (arrows). Scale bar, 10 μm. Images of representative tumour tissue sections from H MIA PaCa-2 xenografts and I the KPCY GEMM stained by IF showing co-localisation of ITGB1 and ILK (arrows). Scale bar, 20 μm. Images of representative primary tumours and liver metastases from J MIA PaCa-2 PDAC xenografts and K the KPCY GEMM stained by IF showing co-localisation of CCDC25 and ILK (arrows). Scale bar, 20 μm. L Immunoblots showing co-immunoprecipitation (co-IP) of ITGB1, ILK and CCDC25 from MIA PaCa-2 human PDAC cells cultured with 5 µg/mL NETs for 24 h. The graph of ILK:ITGB1 ratio is shown to the right. Immunoblots showing co-IP of CCDC25 and ILK from M MIA PaCa-2 human PDAC cells and N KPCY mouse PDAC cells cultured with 5 µg/mL NETs for 24 h. Quantification of band intensities is reported below the blots ( L – N ).

Article Snippet: , Integrin B1 (D2E5) rabbit mAb , CST , 9699 , AB_11178800 , 1:1000.

Techniques: Staining, Immunofluorescence, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Cell Culture

Journal: Pathology

Article Title: A high-impact FN1 variant correlates with fibronectin-mediated glomerulopathy via decreased binding to collagen type IV.

doi: 10.1016/j.pathol.2022.10.016

Figure Lengend Snippet: Fig. 4 Fibronectin variant gain-of-function status was accompanied by integrin b1 overexpression. (A) Immunohistochemistry staining showed that integrin b1 expression was significantly increased in kidney tissue. (B) Western blotting showed integrin b1 expression after HeLa transfection with recombinant plasmids. (C) Immunofluorescence showed integrin b1 overexpression was generated by the fibronectin variant in in vitro transient transfections with plasmids expressing wild-type fibronectin and variant fibronectin. HC, healthy control. Each group was tested in triplicate, and data are presented as mean ± standard deviation. ns, not significant; *p<0.05; ***p<0.001.

Article Snippet: The antibodies used were: anti-fibronectin antibody (1:200, ab2413; Abcam, UK), anti-podocin antibody (1:200, PA5-79757; Invitrogen, USA), HA Epitope Tag antibody (2–2.2.14) (PLA 1:100, 26183; Invitrogen), antiHA Tag antibody (1:500, ab236632; Abcam), anti-integrin b1 antibody (1:100, #34971; Cell Signaling Technology, USA), anti-Col4A3 antibody (1:25, #7076; Chondrex, USA), anti-Col4A4 antibody (1:25, #7073; Chondrex), goat polyclonal secondary antibody to mouse Alexa Fluor 488 (1:400, ab150113; Abcam), goat polyclonal secondary antibody to rabbit Alexa Fluor 555 (1:400, ab150078; Abcam), goat polyclonal secondary antibody to rabbit Alexa fluor 647 (1:400, ab150079; Abcam) and goat anti-rat Alexa Fluor 568 (1:400, ab175473; Abcam), DAPI (1:1000, C1002; Beyotime, China).

Techniques: Variant Assay, Over Expression, Immunohistochemistry, Staining, Expressing, Western Blot, Transfection, Recombinant, Generated, In Vitro, Control, Standard Deviation