ERα localization to lysosomes.

<p>(A) Growing MCF-7 cells were co-stained with anti-ERα Sp-1 and D-12 antibodies. (B) Growing pc DNA flag ERα -transfected HeLa cells were co-stained with anti-ERα HC-20 and flag antibodies. (C) Western blot analysis of ERα cellular levels in growing MCF-7 cells treated with cycloheximide (CHX - 1 mg/ml) for 24 hrs with both anti-ERα Sp-1 and D-12 antibodies. Loading control was done by evaluating vinculin expression in the same filter. (D) Anti-ERα Sp-1 staining of MCF-7 cells treated for with cycloheximide (CHX - 1 mg/ml) for 24 hrs. MCF-7 cells were co-stained with anti-ERα Sp-1 antibody together with either LAMP-2 antibody (E) or lysotracker (F) both in the presence and in the absence of E2 (10 nM–2 hrs). pc DNA flag ERα (Nessi)-transfected HeLa cells were co-stained with anti-ERα HC-20 antibody together with either LAMP-2 antibody (G) or lysotracker (H) both in the presence and in the absence of E2 (10 nM–2 hrs). Figures show one unique confocal plane. All co-staining procedures were described in details in the Material and Methods section.