Anti wnt7a

The relative protein abundances in kidneys and HK-2 cells were analyzed using western blotting, as described previously (Xu et al., 2012 (link)). The following primary antibodies were used: anti-α-SMA (1:1,000, Sigma), anti-E-cadherin (1:1,000, Cell Signaling Technology), anti-vimentin (1:1,000, Santa Cruz Biotechnology), anti-collagen I (1:1,000, Abcam), anti-collagen IV (1:1,000, Abcam), anti-Wnt1 (1:100, Abcam), anti-Wnt4 (1:150, Santa Cruz Biotechnology), anti-Wnt3 (1:500, Abcam), anti-Wnt2b (1:700, Abcam), anti-Wnt7a (1:300, Abcam), anti-β-catenin (1:1,000, Cell Signaling Technology), anti-Snail 1 (1:1,000, Santa Cruz Biotechnology), anti-PAI-1 (1:1,000, Santa Cruz Biotechnology), and anti-NF-κB p65 (1:1,000, Cell Signal). Horseradish peroxidase-conjugated secondary antibodies (anti-rabbit or anti-mouse or anti-goat IgG, 1:5,000, Jackson ImmunoResearch) were used according to the manufacturer’s instructions. Image analysis software (Image J, National Institutes of Health) was used to determine the gray value of all bands.

Cultured cells were lysed in RIPA buffer, and lysates were analyzed using a standard western blot procedure. GAPDH was used as the loading control. PVDF membranes were incubated with anti-GAPDH (1 : 5000), anti-LRP6 (1 : 1000), anti-β-catenin (1 : 1000), anti-Wnt3A (1 : 1000), anti-Wnt7A (1 : 1000), anti-Noggin (1 : 1000), or anti-Wnt5A (1 : 1000) antibodies (Abcam, USA) overnight at 4°C and then washed and incubated for 1 h with secondary antibodies (Abcam, USA).