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

Journal: Molecular oncology

Article Title: Ubiquitin-specific peptidase 39 promotes human glioma cells migration and invasion by facilitating ADAM9 mRNA maturation.

doi: 10.1002/1878-0261.12958

Figure Lengend Snippet: Fig. 7. Knockdown expression of USP39 inhibits the growth and invasion of U251 cells in vivo. The U251 cells expressing USP39 shRNA or negative control shRNA were intracranially implanted into BALB/c nude mice. (A, B) Hematoxylin-eosin (HE) staining of orthotopic xenografts derived from the indicated U251-shUSP39 cells and control in nude mice (n = 3/group). Representative images are shown. Scale bar: 2000, 20 or 100 lm (as indicated in the picture). (C) Kaplan–Meier analysis of survival for tumor-bearing mice implanted with U251-shUSP39 cells and control (n = 6/group). Log-rank test: P < 0.001. (D) Western blotting analysis of USP39, ADAM9 and integrin b1 protein levels in tumor tissues. (E) IHC staining of USP39, ADAM9 and integrin b1 levels in xenograft sections from U251-shNC and U251- shUSP39 groups. Representative images are shown. Scale bar: 20 or 100 lm (as indicated in the picture). Representative data are from three independent experiments.

Article Snippet: The primary antibodies were used as follows: ADAM9 (#ab186833; Abcam, Cambridge, MA, USA), USP39 (#ab131244; Abcam), integrin b1 (#34971; CST, Danvers, MA, USA), GAPDH (#60004-1-Ig; Proteintech, Wuhan, China), b-Actin (#AA128; Hua An Biotechnology, Hangzhou, China).

Techniques: Knockdown, Expressing, In Vivo, shRNA, Negative Control, Staining, Derivative Assay, Control, Western Blot, Immunohistochemistry

Journal: Molecular oncology

Article Title: Ubiquitin-specific peptidase 39 promotes human glioma cells migration and invasion by facilitating ADAM9 mRNA maturation.

doi: 10.1002/1878-0261.12958

Figure Lengend Snippet: Fig. 8. Overexpression of USP39 promotes the growth and invasion of U87 cells in vivo. (A, B) Representative images of HE staining of orthotopic xenografts obtained from the indicated U87-ov- USP39 cells and control in nude mice (n = 3/group). Scale bar: 2000, 20 or 100 lm (as indicated in the picture). (C) Kaplan–Meier analysis of survival for tumor-bearing mice implanted with U87- ov-USP39 cells and controls (n = 6/group). Log-rank test: P < 0.05. (D) Western blotting analysis of USP39, ADAM9 and integrin b1 protein levels in tumor tissues. (E) IHC staining of USP39, ADAM9 and integrin b1 levels in xenograft sections from U87-ov-Con and U87-ov-USP39 groups. Representative images are shown. Scale bar: 20 or 100 lm (as indicated in the picture). Representative data are from three independent experiments.

Article Snippet: The primary antibodies were used as follows: ADAM9 (#ab186833; Abcam, Cambridge, MA, USA), USP39 (#ab131244; Abcam), integrin b1 (#34971; CST, Danvers, MA, USA), GAPDH (#60004-1-Ig; Proteintech, Wuhan, China), b-Actin (#AA128; Hua An Biotechnology, Hangzhou, China).

Techniques: Over Expression, In Vivo, Staining, Control, Western Blot, Immunohistochemistry