Figure 5: Kctd10 directly binds to Tbx5/5a to moderate its transcriptional activity.

(a,b) Luciferase reporter assays of the G5E1b and the 0.7-kb Anf promoter fragment. Numbers indicate the quantity (nanograms) of each corresponding plasmid. At least three independent experiments were performed for each assay, and each in quadruplicate (mean±s.e.m.; n=4). Asterisks represent the statistical significance as determined using a Student’s t-test (*P<0.05; **P<0.01). (c) Co-IP of endogenous TBX5 and endogenous KCTD10 in HeLa cells. (d) Pull-down assay of His-tagged Tbx5a by GST fusion Kctd10 protein bound to GSH-agarose beads and subsequent detection by western with anti-His antibody. The expression of GST and GST fusion protein was stained by Coomassie brilliant blue. (e) Immunofluorescent detection of HA-Tbx5a and Kctd10-GFP in 293T cells. Nuclei are counterstained by DAPI. Scale bar=10 μm.