Skip to main content
. 2008 Jun;19(6):2588–2596. doi: 10.1091/mbc.E08-02-0187

Figure 4.

Figure 4.

LRP6 is implicated in RSpo-mediated β-catenin signaling. (A) HEK293 cells stably expressing 16-TCF reporter were transfected with either wt LRP6 or LRP6ΔC expression constructs, treated with indicated RSpo proteins (200 ng/ml), and the luciferase reporter activity was determined 20 h after the incubation. Empty vector was used as a transfection control. (B) Regular HEK293 cells were treated with RSpo proteins (200 ng/ml), Wnt3A (200 ng/ml), or Wnt3A (200 ng/ml), and individual RSpo protein (200 ng/ml) together for 4 h. Total cell lysates were prepared and resolved by SDS-PAGE. Phosphorylation of endogenous LRP6 was detected by immunoblotting using anti-phospho-LRP6 antibodies. (C) HEK293 cells were stably transfected with a LRP6-FLAG expression construct and treated with Wnt3A (200 ng/ml), RSpo proteins alone (200 ng/ml) or Wnt3A and RSpo together for 4 h. The total cell lysates were prepared and resolved by SDS-PAGE followed by immunoblotting using anti-phospho-LRP6 antibodies. (D) Stable HEK293 A6 cells were treated with Wnt3A (250 ng/ml) or RSpo protein alone (125 ng/ml) or in combination with recombinant DKK1 protein (400 ng/ml). The luciferase reporter activity was determined 20 h after incubation.