Figure 3. AREG mediates the effect of TAZ. (A) AREG-neutralizing antibody blocks TAZ^S89A and TAZ^4SA-induced EGF-independent 3D growth. Vector control and TAZ-expressing cells were cultured in 3D assay in the absence of EGF for 12 d, together with a neutralizing antibody against AREG. Normal goat IgG is used as a control. Representative images are shown. (Scale bar, 100 µm) (B) Efficient knockdown of AREG by shRNA in vector control or TAZ^S89A and TAZ^4SA-transduced MCF10A cells, as revealed by immnoblot. β-Actin was used as a loading control. (C) Knockdown of AREG abolishes the 3D phenotype induced by TAZ^S89A and TAZ^4SA in absence of EGF (Scale bar, 100 µm). (D) Knockdown of AREG reduces the TAZ^S89A- and TAZ^4SA-induced cell migration.