Restore western blot stripping solution

To assess PBX1 expression levels in tissues, we collected cells from endometrial epithelial cells (EME), ovarian surface epithelial cells (OSE), fallopian tube epithelial cells (FTE), ovarian clear cell carcinoma (CCC), and low-and high-grade serous carcinoma (LGSC and HGSC). After washing, cells were lysed with lysis buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 1% NP40) supplemented with protease inhibitor cocktail (Thermo Scientific). Cell lysates were separated by SDS-PAGE and transferred onto a PVDF membrane by a semi-dry transfer (Bio-Rad). After blocking with 5% non-fat dry milk in TBST (20 mM Tris-HCl, 0.5 M NaCl, 0.1% Tween 20), samples were incubated overnight with anti-PBX1 antibody and subsequently incubated in secondary antibody (Jackson Laboratories, West Grove, PA). After developing with ECL solution (Amersham), PVDF membranes were stripped with Restore Western Blot Stripping Solution (Thermo Scientific) and then re-blotted with anti-GAPDH antibody for the loading control. To calculate PBX1 protein expression levels, the intensity of PBX1 and GAPDH bands were determined using ChemiDoc XRS (Bio-Rad), and levels were calculated using the following formula: (IntPBX1/IntGAPDH)/lowest(IntPBX1/IntGAPDH).