Long-range Wnt-5A inhibits the ATF2-Luc reporter.

<p>A) For studying long-range signaling, Xwnt5A-EGFP transfected HEK293 cells seeded on thinCerts TC chambers were transferred on cells transfected with the ATF2-Luc reporter and cultivated for additional 24 hours. To investigate paracrine signaling, cells transfected with Xwnt5A-EGFP were co-cultured with cells transfected with the ATF2-Luc reporter for 24 hours. To analyze both autocrine and paracrine signaling, the reporter and the Wnt ligand were cotransfected. B) Activation of the non-canonical Wnt reporter ATF2-Luc by co-transfected Wnt8, Wnt11 and Wnt5A. C) Activation of the non-canonical Wnt reporter ATF2-Luc by long-range Wnt8, Wnt11 and Wnt5A in a two-chamber assay. D) Xwnt5A-EGFP triggered ATF2-Luc activation depends on endocytosis. Wnt5A induced ATF2-Luc activation (cotransfection) was inhibited by addition of 5 µg/ml chlorpromazine (Chlo) and 150 µM genistein (Gen) 24 h before the measurement. Given are the mean values ± standard errors and the p-values according to Student's t-test, ns: not significant, n gives the number of transfections.