Anti her2

In-cell ELISA was performed using the In-Cell ELISA Kit (Abcam, France) according to the manufacturer’s instructions and after a fixation step performed with 4% of paraformaldehyde solution (10 min at room temperature). Primary antibodies were incubated overnight at 4°C. Adequate HRP-conjugated secondary antibodies were incubated for 1 h at room temperature. Detection was performed at 450 nm.
After the washes, cells in each well were incubated with 1X Janus Green Stain for 5 min at room temperature, according to the manufacturer’s instructions. Data were expressed in normalized unit, according to the following calculation: (HRPsignal ‘minus’ HRPsignal in absence of primary antibody)/(Janus Green signal ‘minus’ Janus Green signal in absence of cells).
Antibodies used were anti-TET1 (sc163446, Santa Cruz, France), anti-TET2 (sc398535, Santa Cruz), anti-TET3 (sc139186, Santa Cruz), anti-ERα (sc8002, Santa Cruz), anti-PR (sc130071, Santa Cruz), and anti-HER2 (sc-393712, Santa Cruz).

OVCAR3 and A2780 cell lines were processed for protein isolation and Western blotting using standard procedures, as described previously [19 (link)]. The following primary antibodies were used: anti-hPaf1/PD2, anti-Leo1, anti-Parafibromin, anti-Ctr9 (Bethyl Laboratories, Montgomery, TX, USA); anti-OCT3/4, anti-SOX-2, anti-CD24, anti-ESA, anti-SHH, anti-HER2 (Santa Cruz Biotechnology, Dallas, TX, USA); anti-CD44 (Cell signalling Technology, Danvers, MA, USA); anti-CD133 (Abnova, Walnut, CA, USA); anti–β-Catenin (Sigma); and anti-Lgr5, anti-SOX-9 (Abcam, Cambridge, MA, USA) overnight at 4°C. β-actin was used as a loading control. The band intensity was quantified using ImageJ and the normalization was performed as described previously [18 (link)].