Journal: International Journal of Molecular Sciences

Article Title: Change in Renal Glomerular Collagens and Glomerular Filtration Barrier-Related Proteins in a Dextran Sulfate Sodium-Induced Colitis Mouse Model

doi: 10.3390/ijms20061458

Figure Lengend Snippet: Changes in glomerular collagens in mice after DSS administration. Immunofluorescent microscopy ( A ) and Western blot analysis of protein expression ( B ) for type IV collagen (COL IV; A-a, A-a’; B-a; 160–190 kDa), type I collagen (COL I; A-b, A-b’; B-b; 150 kDa), and type V collagen (COL V; A-c, A-c’; B-c; 220 kDa) were conducted for control and DSS-colitis mice. Representative bands ( B , left) and relative band intensity ratios were analyzed ( B , right). ( C ) Illustration of glomerular collagens changes in this study. All values are means ± SEM ( n = 6); * p < 0.05 and ** p < 0.01 vs. control. Scale bars = 10 μm. Abbreviations: GBM, glomerular basement membrane; BC, Bowman’s capsule.

Article Snippet: Sections of 5-μm thickness were cut by a cryostat (CM3050; Leica, Wetzlar, Germany); then incubated with primary antibodies against type I collagen (Acris Antibodies, Germany), type IV collagen, type V collagen, synaptopodin, VE-cadherin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), nephrin, and podocalyxin (R&D Systems, Minneapolis, MN, USA), respectively, at 4 °C overnight, and followed by secondary antibodies conjugated to Alexa Fluor ® 488 or 568 (Invitrogen, Carlsbad, CA, USA); double-stained with rhodamine-conjugated phalloidin (Life Technologies, Gaithersburg, MD, USA) for F-actin; and finally submerged in fluoroshield mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI) (Abcam, Cambridge, UK).

Techniques: Microscopy, Western Blot, Expressing, Membrane