Figure 6. WNT5A induces the phosphorylation of SMAD2 in a TGFβ receptor-dependent fashion.
A. Basal and luminal mammary epithelial cells were purified from pre-neoplastic mammary gland of 5-month-old ErbB2/WNT5A+/− mice and cultured in 24 well plates for 72 hrs. Cells were either mock-treated, treated with WNT5A (100ng/ml), WNT5A+SB-431542 (5 µM), or SB-431542 alone for 1 hr. B. MCF10A cells were transiently transfected with control siRNA (siCON), or two independent siRNAs to TGFβR1 (si-1, si-3). 96 hrs later, cells were either treated with WNT5A (100ng/ml) or TGFβ (5 ng/ml) for 1 hr. For A–B, cell lysates were collected, separated by SDS-PAGE and immunoblotted with the indicated antibodies (n=2–3). Band densitometry was measured by Image J.