FIG 9.

KDM2B induces AP-1 transcriptional activity through its F-box domain. (A) Identification of host signaling pathways that are regulated by KDM2B using luciferase reporter assay. HEK293T cells were cotransfected with 3×FLAG-KDM2B WT or CXXCm along with luciferase reporter plasmids whose promoters are responsive to the signaling pathways indicated on the graph. The fold change was calculated by comparing KDM2B-induced luciferase activity to the basal activity of the luciferase reporter plasmids alone. (B) Immunoblotting analysis of transfected WT and KDM2B mutants. Tubulin was used as a loading control. (C) KDM2B mutants were tested in the AP-1 luciferase reporter assay as described for panel A. (D) Expression of 3×FLAG-LRR reduced the KDM2B-induced AP-1 activity. HEK293T cells were cotransfected with AP-1 luciferase reporter plasmid and 3×FLAG-KDM2B WT along with an increasing amount of 3×FLAG-LRR plasmid (50 to 600 ng). The y axis shows luciferase light units. t tests were performed between samples with and without KDM2B or CXXCm (A) and between vector and KDM2B samples (C), and P < 0.05 (*) was considered statistically significant.