Compound 3289–8625 suppressed β-catenin protein level, thus regulating Wnt activity in HRMECs. HRMECs were treated with LCM (A), WCM (B), or WCM in presence of compound 3289-8625 (50 μM) (C), for 24 hours. Five random fields were examined on the slide. The green channel of phalloidin staining represents the HRMECs cytoskeleton. Immunofluorescence analysis of total β-catenin protein levels, as indicated by the red channel in HRMECs, shows that WCM enhanced the translocation of β-catenin protein levels into the nucleus. Moreover, compound 3289-8625 can inhibit Wnt signaling activity in HRMECs. HRMEC, human retinal microvascular endothelial cell; LCM, L-cell control conditional medium; WCM, Wnt-3a conditional medium