Fig. 3: Kaempferol targets the kinase domain of FGFR3. | Cell Death & Disease

Fig. 3: Kaempferol targets the kinase domain of FGFR3.

From: Kaempferol targeting on the fibroblast growth factor receptor 3-ribosomal S6 kinase 2 signaling axis prevents the development of rheumatoid arthritis

Fig. 3

a Docking scores calculated by Protein Preparation Wizard in Maestro v9.2 using kaempferol 3D structures (CID 5282102) and crystal structures of indicated RTKs obtained from the Protein Data Bank. b The lowest docking score of kaempferol to the active pocket of FGFR3 represents the formation of hydrogen bonds with Lys508, Glu555, and Ala558, as indicated. c Kaempferol competition assay. Binding of FGFR3 kinase domain to ATP-conjugated agarose beads was competed with the indicated doses of kaempferol. The graph indicates fold change of band intensity from three independent western blots of the FGFR3 pull-down assay. Data were obtained from three independent experiments, and values are represented as means ± SEM. *p < 0.01; **p < 0.001. d, e Kaempferol binding to FGFR3 was confirmed by a CNBr-kaempferol pull-down assay using a commercial FGFR3 kinase domain (c) and a membrane fraction of MH7A cells (d). The western blot is representative of three independent experiments. f, g The effects of kaempferol on bFGF-induced FGFR3 phosphorylation at Tyr724 was analyzed by western blotting (f) and immunocytofluorescence (g) using indicated p-FGFR3 specific antibodies in MH7A cells. The band intensity of western blot (f) was measured using Image J (Ver. 1.6), and each indicated area (g) in the immunocytofluorescence confocal image (×400) was magnified (×630). Scale bars, 20 μm

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