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Supplementary Figure 3. TGF-β regulates estrogen signaling and EGFR expression in BT474 cells. (A) Differential mRNA expression of ER-a and ER-b in quiescent BT474 cells. (B) BT474 cells were stimulated with TGF-β1 (5 ng/mL) for 0-96 h as indicated. Afterward, total RNA was harvested and subjected to real-time PCR to monitor differences in ER-a expression. (C) BT474 cells were transiently transfected overnight with p3TP-, pSBE-, or pERE-luciferase and pCMV-bgal cDNAs prior to their stimulation for 24 h with TGF-β1 (5 ng/mL). Afterward, luciferase and bgal activities contained in detergent-solubilized cell extracts were measured. Data are the mean fold-changes (± SE; n = 3;*P < 0.05; Student's t-test). TGF: transforming growth factor; EGFR: epidermal growth factor receptor; SE: standard error