Anti gpr31

Proteins were electrophoresed by SDS/PAGE (12% or 10% gel) and the blots were incubated overnight with primary antibody. The following primary antibodies were used: anti-ALOX12 (Santa-Cruz, sc-365,194), anti-E-Cadherin (Cell signal Technology, #14472), anti-N-Cadherin (Cell signal Technology, #13116), anti-Vimentin (Cell signal Technology, #5741), anti-Snail (Cell signal Technology, #3879), anti-Slug (Cell signal Technology, #9585), anti-MMP2 (Abcam, ab37150), anti-MMP7 (Abcam, ab5706), anti-MMP9 (Abcam, ab38898), anti-MMP13 (Abcam, ab39012), anti-GPR31 (Abcam, ab75579), anti-PI3K (Cell signal Technology, #4249), anti-AKT (Abcam, ab179463), anti-AKT (phospho T308, Abcam, ab38449), anti-NF-κB (Abcam, ab16502), anti-NF-κB (phospho S536, Abcam, ab86299), anti-GAPDH (Abcam, ab181602).

Proteins were electrophoresed by SDS/PAGE and blots were incubated overnight with primary antibody. The following antibodies were used: anti-GPR31 (Abcam, ab75579), anti-ATPase Na⁺/K⁺ β2 (Bioss, bs-1152R), anti-Wnt3a (Cell signaling Technology (CST), #2391), anti-Wnt5a (CST, #2530), anti-pGSK-3β (phospho Ser9, CST, #5558), anti-JNK (phospho Thr183/Tyr185, CST, #4671), anti-β-catenin (CST, #8480), anti-HA tag (CST, #5017), anti-flag tag (CST, #14,793), anti-USP24 (Proteintech, 13,126–1-AP), anti-FGGY (Abnova, ABN-H00055277), anti-β-Actin (CST, #4967), and anti-GAPDH (Abcam, ab181602). After incubated with HRP-conjugated α-rabbit or α-mouse secondary antibodies for 1 h, protein bands were detected with chemiluminescence substrate (Perkin Elmer) using the ChemiDoc Imaging System (Bio-Rad).

All procedures were performed as described above [89 (link)]. The antibodies are as follows: anti-FGGY (Abcam), anti-12-LOX (Abcam), anti-15-LOX (Abcam), anti-GPR31 (Abcam), and a biotinylated secondary goat anti-mouse IgG antibody (Santa Cruz), labeled with HRP using a DAB staining kit (Maixin Biotechnology) according to the manufacturer’s instructions. For negative controls, IgG1 was used. Positively stained cells were counted in 5 fields at 200 × magnification.