Whole cell extracts were prepared using lysis buffer containing 20 mM Tris, 150 mM NaCl, 1% NP-40, 1% SDS, 5% glycerol, and the Complete Mini protease inhibitor cocktail tablet (Roche). Cytoplasmic and nuclear fractions were isolated using CE buffer (HEPES [10 mM] pH 7.9, KCI [10 mM], EDTA [0.1 mM], NP-40 0.3% (added just before use) and 1X protease inhibitors (added just before use)) and NE buffer (HEPES [20 mM] pH 7.9, NaCl [0.4 M], EDTA [1 mM], glycerol 25%, 1X protease inhibitors (added just before use)) exactly as detailed previously [19 (link)]. Proteins were separated on 4–12% SDS-PAGE gels, and transferred to nitrocellulose membranes. The membrane was dried for 1 h to bind proteins tightly to the membrane and blocked with Intercept (PBS) Blocking Buffers (LI-COR) before incubation with primary antibody. Western blotting used antibodies against ERβ isoforms (MC-10, Thermo Fisher 14-9336-82), ERβ5 (Bio-Rad, Clone 5/25, MCA4676GA), survivin (Cell Signaling Technologies), E-cadherin (Cell Signaling Technologies), and β-actin (Sigma-Aldrich) as an internal loading control. Both IRDye 800 CW goat anti-rabbit secondary antibody (LI-COR, 926-32211) and IRDye 680 CW goat anti-mouse secondary antibody (LI-COR, 926-68070) were diluted (1:5000) for incubation with the blots. Quantification of protein bands used the LI-COR Odyssey CLx Imaging System and LI-COR analysis software.