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E2 transiently decreases TGFβ-induced Smad signaling in bone-tropic ER+ breast cancer cells. Smad2/3 expression and TGFβ-stimulated (5 ng/mL for 1 h) Smad2 and Smad3 phosphorylation in (A) MCF-7 and (B) MCF-7 BMET-derived 43-4M cells maintained in E2-deplete media for 4 days prior to stimulation with E2 (10−8 M) vs. E2-deplete media for the indicated times prior to addition of TGFβ (5 ng/mL) for one hour. Note, early (0–5 h) vs. late times (15–23 h) were run on two separate blots.
