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. 2000 Mar 7;97(7):3130–3135. doi: 10.1073/pnas.050015697

Figure 3.

Figure 3

Immunofluorescence analysis of RIZ protein in MCF-7 cells. (Upper) Cells were incubated without hormones (A and G), with 50 nM estradiol (B, C, and H), with 10 nM estradiol and 10 μm ICI182,780 (D), with 1 μM all-trans-retinoic acid (E), or with 100 ng/ml 12-O-tetradecanoylphorbol-13-acetate (F). (A, B, and D–F) Staining with antibody to RIZ protein RZ1802. (C) Permeabilization with Triton X-100 followed by staining with antibody to RIZ protein RZ1802. (G and H) Staining with antibody to proliferating cell nuclear antigen. (Lower) Cells were grown in Petri dishes in hormone-free medium and, subsequently, were incubated with 50 nM estradiol (E2), with 10 nM estradiol and 10 μm ICI182,780 (ICI), with 1 μM all-trans-retinoic acid (ATRA), or with 100 ng/ml 12-O-tetradecanoylphorbol-13-acetate (TPA) or kept in hormone free-medium (none). Harvested cells were permeabilized and processed for flow cytometry as described in Materials and Methods. Results were measured as number of molecules of equivalent soluble fluorochrome (MESF).