Skip to main content
. 2016 Sep 21;7:12887. doi: 10.1038/ncomms12887

Figure 1. Parafibromin transactivates Hedgehog signalling in a tyrosine dephosphorylation-dependent manner.

Figure 1

(a) Parafibromin knockdown results in reduced Hh signal activity. Hh-responsive luciferase reporter assays were performed in HEK293T cells. Total cell lysates (TCLs) were subjected to immunoblotting (n=3, mean±s.d. **P<0.01; ANOVA with Bonferroni's post hoc test). (b) Parafibromin interacts with Gli1. TCLs prepared from SKES cells were sequentially immunoprecipitated (IP) with an anti-parafibromin antibody and immunoblotted. (c) Phospho-resistant (PR)-parafibromin shows increased ability to activate Hh signalling. Hh-responsive luciferase reporter assays were performed in HEK293T cells (n=3, mean±s.d. **P<0.01; ANOVA with Bonferroni's post hoc test). (d) SHP2 expression promotes Hh signalling. Hh-responsive luciferase reporter assays were performed in A549 cells (n=3, mean±s.d. **P<0.01; a two-tailed unpaired Student's t-test). (e,f) PR-parafibromin shows increased binding to β-catenin and Gli1. HEK293T cells were transfected with the indicated vectors. TCLs were sequentially immunoprecipitated (IP) and immunoblotted. (g,h) Non-phosphorylated parafibromin specifically interacts with β-catenin and Gli1. HEK293T cells were transfected with the indicated vectors. TCLs were immunoprecipitated with the indicated antibodies and immunoprecipitates (IP) were incubated with recombinant phosphorylated/non-phosphorylated parafibromin. The reaction mixtures were subjected to immunoblotting.