Journal: Cells

Article Title: Pharmacological Blockade of the Adenosine A 2B Receptor Is Protective of Proteinuria in Diabetic Rats, through Affecting Focal Adhesion Kinase Activation and the Adhesion Dynamics of Podocytes

doi: 10.3390/cells13100846

Figure Lengend Snippet: Modulation of A 2B AR affects the adhesion and migration of podocytes. ( A ). Representative indirect immunofluorescence images of podocytes at a spreading of 60 min and treatments using adenosine (10 μM) or a combination of the A 2B AR antagonist MRS1754 (50 nM). The phalloidin staining of actin filaments is pseudo colored in green; Y397-phosphorylated FAK is in red; nuclear staining with DAPI is in blue. n = 3. Bar indicates 100 µm. ( B ). Representative wound healing assay images. Monolayer cultures of differentiated podocytes were exposed to adenosine and in combination with the A 2B AR antagonist MRS1754. Light microscopy images were captured at wounding and following 6 and 24 h. Bar indicates 300 µm. ( C ). Quantification of wound healing percentage from images of treated podocytes in ( B ). The graph represents means ± SEM. Five fields were analyzed by each condition. Four independent experiments were achieved. ( D ). Representative images of podocyte transmigration tests through Boyden’s chambers following treatments with adenosine and in combination with the A 2B AR antagonist MRS1754. Bar indicates 300 µm. ( E ). Quantitative analysis of transmigrated cells from experiments in ( D ). The graph depicts means ± SEM of the number of cells per field, 5 fields analyzed in each condition. n = 3. p values are shown on brackets in graphs ( C , D ) obtained from ANOVA with Fisher’s least significant difference (LSD) post hoc test. *, p < 0.05; n.s., no statistical difference.

Article Snippet: Data were tabulated and analyzed by ANOVA with (LSD) post hoc test using GraphPad Prism 9.0.

Techniques: Migration, Immunofluorescence, Staining, Wound Healing Assay, Light Microscopy, Transmigration Assay