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