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. Author manuscript; available in PMC: 2012 Mar 1.
Published in final edited form as: Cancer Res. 2011 Jan 6;71(5):1658–1668. doi: 10.1158/0008-5472.CAN-10-1899

Figure 2. Effects of EGF and calcium on ERα activity.

Figure 2

A. Effects of EGF and calcium on ERα activity in MCF-7 cells. Cells were treated for 1 hour with estradiol (1nM), calcium (5mM), or EGF (150ng/ml) and the occupancy of ERα on the pS2 promoter was examined using a chromatin immunoprecipitation assay.

B. Effects of calcium on growth. Cells treated with estradiol (1nM) and calcium (0.25, 0.5, 1.0, and 2.0mM) on day 0 or calcium (0.25, 0.5, and 1.0mM) on days 0, 1, 2 and 3 in the presence and absence of the antiestrogen ICI-182,780 (500nM). Cell number was measured on day 4 (mean±SD; three independent experiments). *, p < 0.05; **; p < 0.005; ***, p < 0.0005. a, compared to control; b, compared to treatment.

C. Effects of EGF and calcium on ERα target genes in MCF-7/LCC2 cells. MCF-7/LCC2 cells were treated for 24 hours with estradiol (1nM), EGF (150ng/ml), or calcium (5mM) in the presence and absence of the antiestrogen ICI-182,780 (500nM) or the intracellular calcium chelator BAPTA-AM (1uM). The amount of PgR mRNA was measured using a real time RT-PCR assay and normalized to the amount of 18S ribosomal RNA. Data are expressed as percent control (mean±SD; two independent experiments done in duplicate). *, p < 0.05; **; p < 0.005; ***, p < 0.0005. a, compared to control; b, compared to treatment