Alternative isoforms of KDM2A and KDM2B lysine demethylases negatively regulate canonical Wnt signaling
Fig 2
KDM2A-SF and KDM2B-SF repress the canonical Wnt signaling luciferase reporter.
A. The wild type TCF/LEF reporter (TOP5) contains five TCF/LEF consensus binding sites (CTTTGAT) that drive the expression of the luciferase gene. The FOP5 construct contains five mutant TCF/LEF binding sites (CTTTGCC) instead and serves as the background activity control. B. Both KDM2A-LF and KDM2A-SF strongly repressed the TOP5 reporter activated with BIO. The reporter activity is expressed as the fold change ratio between the normalized luciferase signal of TOP5 and FOP5. C. KDM2B-SF, but not KDM2B-LF, also strongly repressed the activated reporter. D. Repression of the TOP5 reporter by KDM2A-LF is not dependent on the activity of its JmjC demethylase domain, but on the CXXC DNA binding domain, PHD domain and HP1a interaction motif. The HP1a motif of KDM2A-SF is not necessary for the repressive effect. E. KDM2B-LF was not able to repress the activated TOP5 reporter, whereas KDM2B-SF strongly repressed it in a DNA binding domain dependent manner independently of the PHD domain. (*p < 0.05; **p < 0.01).