Anti wnt7a antibody

RIPA lysis buffer (Thermo Fisher Scientific, USA) containing protease and phosphatase inhibitors was used to lyse tissues. Protein lysates were separated by SDS-PAGE gels (Thermo Fisher Scientific, USA), blotted onto PVDF membrane (Roche, Switzerland) for analysis and incubated at 4°C overnight with the following primary antibodies: anti-WNT2B antibody (1:1000 dilution; Bioss, China), anti-WNT7A antibody (1:1000 dilution; Bioss, China), and anti-GAPDH antibody (1:2000 dilution; Cell Signaling Technology, USA). The results of the western blot analyses were performed with Image J software.

Immunohistochemistry was performed in 20 NSCLC tissues and adjacent normal tissues. Paraffin-embedded sections were stained to determine the expression level of proteins. Sections were incubated overnight at 4°C with anti-WNT2B antibody (1:100 dilution; Bioss, China), or anti-WNT7A antibody (1:100 dilution; Bioss, China). After washing with phosphate-buffered saline (PBS), the slides were incubated with a goat anti-rabbit IgG secondary antibody conjugated to fluorescein isothiocyanate (ZSDB-BIO, China) for 30 mins. They were washed with PBS and then incubated with an antifade reagent (Invitrogen, USA). Finally, staining was observed using an Olympus CX41 fluorescence microscope (Olympus, Japan). The results of the analyses were performed with Image J software.