Fig. 7.

Effects of expression of Smad2/3/4 and constitutively active TGF-β type I receptor on transcription from Zp. EBV-negative AGS cells in 24-well plates were cotransfected as indicated with 80 ng of the firefly luciferase reporter, pZp-Luc-WT (i), 100 ng of pCS2-FLAG-Smad2 and/or pCS2-FLAG-Smad3 as indicated (ii), 100 ng of pCMV6-XL5 Smad4 (iii), and 50 ng of pcDNA3-ALK5 T204D (iv) where indicated by light gray bars. The empty vector, pcDNA3.1, was added to bring the total DNA to 430 ng per well. Cell lysates were prepared 48 h later, and firefly luciferase activities were measured. Data were internally normalized to the amount of protein in each lysate and externally normalized to the basal activity of pZp-Luc-WT. Bars indicate means ± standard deviations (SD) of data from assays performed in quadruplicate.