FGF9 and Wnt Pathway Protein Analysis

Palates of embryos was extracted at E13.5 days. Western blotting was carried out as previously described [21 (link)]. Proteins were transferred to PVDF membranes (Millipore, Billerica, MA, USA), which were incubated overnight at 4 °C with rabbit anti-FGF9 antibody (1:500, ab206408, MA, Abcam), rabbit anti-TCF7L2/TCF4 antibody (1:1000, 13838-1-AP; Proteintech, Wuhan, China), rabbit anti-β-catenin antibody (1:500, ab223075; Abcam), rabbit anti-HAS2 antibody (1:500, ab290669; Abcam), and rabbit anti-GADPH antibody (1:5000, Proteintech, Wuhan, China). Afterwards, these primary antibodies were detected using HRP-conjugated anti-rabbit IgG antibody (Servicebio, Wuhan, China) for 1 h at room temperature. Finally, the protein-specific signals were detected using a Bio-Rad ChemiDoc™ MP Imaging System.