Journal: The Journal of Biological Chemistry

Article Title: Small molecule intervention of actin-binding protein profilin1 reduces tumor angiogenesis in renal cell carcinoma

doi: 10.1016/j.jbc.2025.111033

Figure Lengend Snippet: Discovery of a novel analog of C74 with superior antiangiogenic activity in vitro and in vivo . A and B , docking models of C74 and UP-6 interaction with Pfn1—the binding modes of C74 ( panel A ) and UP-6 ( panel B ) as predicted by GNINA. Blue arrow indicates the altered structure on each compound. The unmodified hydroxypyrazoles are predicted to make an identical network of hydrogen bonds in both compounds while the modified aryl ring can potentially interact with R74 and H119. C and D , representative images ( panel C ; 4x magnification) and quantification ( panel D ; based on 4x field images) of cord formation by HmVECs on Matrigel treated with either 20 μM compound UP6 or equivalent DMSO control. Cord length is determined by the distance between each cord junction, i.e. cord intersections. Each data point represents quantification of cord formation in a single field of observation (two-tailed Student’s t test; ∗∗∗∗, p < 0.0001; n = 3 experiments; the scale bar represents 200 μm). E - F , representative fluorescence images ( panel E ) and quantification ( panel F ) of cord formation by HmVECs (labeled with cell tracker dye) subjected to 10 μM of either C74 or UP6 versus DMSO (control) (One-way ANOVA with Tukey multiple comparison post hoc test, ∗ - p < 0.05; ns – not significant; the scale bar represents 200 μm). G - H , representative images of CD31 immunohistochemistry with hematoxylin counterstaining ( panel G ; 4 × magnification) and quantification ( panel H ; based on 20X field images) of CD31+ cell infiltration in the subcutaneously implanted Matrigel plugs in Balb/C mice coinjected with a single 200 μM dosage of either C74 or UP-6 or DMSO control. Data are summarized from quantification of 10 bilaterally injected plugs (n = 5 mice) per treatment group (One-way ANOVA with Tukey post hoc test, ∗∗, p < 0.01; ∗∗∗∗, p < 0.0001; the scale bar represents 200 μm). Pfn1, profilin1; HmVEC, human microvascular EC; DMSO, dimethyl sulfoxide.

Article Snippet: For testing of anti-angiogenic capability of small molecules, HmVECs were plated on growth factor-reduced matrigel (R&D Systems, 3433-005-01) and treated with various compounds at a 20 μM concentration (unless indicated otherwise) or equivalent concentration of DMSO as described previously ( ).

Techniques: Activity Assay, In Vitro, In Vivo, Binding Assay, Modification, Control, Two Tailed Test, Fluorescence, Labeling, Comparison, Immunohistochemistry, Injection