SETD6 is required for β-catenin transcriptional activity at chromatin.
A, HEK-293T control and SETD6 knock-out cells were transfected with FLAG-PAK4 and with a control plasmid or LRP6 ΔN plasmid to activate the Wnt pathway. After 48 h, chromatin extracts were incubated with FLAG M2 beads and A/G beads conjugated to IgG as a control. Input shows the levels of FLAG-PAK4, β-catenin, SETD6, and H3 in the chromatin fraction. B, quantification of GFP-positive cell numbers in HEK-293T control and SETD6 knock-out cells transfected with TOP-GFP that were activated with LRP6 ΔN or transfected with empty plasmid. Data are from at least three experiments (error bars, S.E.). C, same as in B, except that after transfection, cells were harvested and subjected to Western blot analysis with the indicated antibodies. D, suggested model for how SETD6 regulates Wnt/β-catenin signaling.