Fig. 5

S100A8/A9-RAGE-FAK signaling triggers YAP activity in TNBC. A pS127YAP, YAP, pT183/180-MST1/2 and MST1 immunoblots in MDA-MB231 cells stable overexpressing RAGE and treated for 60 minutes with 100 ng/ml rhS100A8/A9 alone or in combination with 1 μM RAGE antagonist FPS-ZM1. B pS127YAP, YAP, pT183/180-MST1/2 and MST1 immunoblots in MDA-MB231 cells overexpressing RAGE and treated for 60 minutes with 100 ng/ml rhS100A8/A9 alone or in combination with 1 μM FAK inhibitor VS-4718. C YAP nuclear immunofluorescence staining in MDA-MB231 cells overexpressing RAGE and treated for 60 minutes with 100 ng/ml rhS100A8/A9 alone or in combination with 1 μM RAGE antagonist FPS-ZM1 or 1 μM FAK inhibitor VS-4718. D Relative quantification of cells with enhanced YAP nuclear localization. E YAP nuclear and cytoplasmic fractionation in MDA-MB231 cells overexpressing RAGE treated for 60 minutes with 100 ng/ml rhS100A8/A9 alone or in combination with 1 μM RAGE antagonist FPS-ZM1 or 1 μM FAK inhibitor VS-4718, using lamin A/C and ꞵ-actin as nuclear and cytoplasmic control markers, respectively. F YAP/TAZ luciferase reporter assay in MDA-MB231 cells overexpressing RAGE and treated for 6 hours with 100 ng/ml rhS100A8/A9 alone or in combination with 1 μM RAGE antagonist FPS-ZM1 or 1 μM FAK inhibitor VS-4718. Error bars represent mean ± SD. * indicates p-value < 0.05. ꞵ-actin served as loading control for immunoblots. Results shown are representative of three independent experiments performed in triplicate