Figure 6: JMJD1A (KDM3A) stabilizes the GLI1 protein.

(a) Co-immunoprecipitation experiment in Hek293T cells detecting an interaction between endogenous KDM3A and endogenous GLI1. (b) Co-transfection of GFP-tagged GLI1 or GLI2 (full length) and the indicated constructs in Hek293A cells. SUFU was included as positive control for GLI stabilization. Shown are representative results of three independent experiments. (c) Quantification of the results depicted in b. Shown is the mean GLI band intensity normalized to actin of n=3 experiments (±s.d.). (d) Western blot of Hek293A cell lysates after transfection of GLI1-GFP and the indicated constructs. Shown are representative results of three independent experiments. (e) Determination of the half-life of endogenous GLI1 protein in Hek293T cells transfected with empty vector control (pcDNA), KDM3A or its catalytically inactive mutant (KDM^mut). CHX, cycloheximide (100 μg ml⁻¹) for the indicated time periods. Shown are representative results of three independent experiments. (f) Quantification of the results depicted in e. Shown is the mean GLI band intensity normalized to actin of n=3 experiments (±s.d.). (g) Western blot result of co-transfection experiments in Hek293A cells. Shown is a representative result of three independent experiments. *P<0.05; **P<0.005; ***P<0.0005 (Student’s t-test).