The BAC-SMAD2 and BAC-SMAD3 recombinant plasmids used in this study were provided by the Genome Engineering Core Facility of the Institute for Genomics and Systems Biology at the University of Chicago. A BAC containing the gene and endogenous cis control elements was tagged by recombineering to yield the Localization and Affinity Purification(LAP) tag at the C-terminus [18]. Smad2 was tagged in CH17-5E15BAC (BAC-SMAD2) while Smad3 was tagged in CH17-187G10BAC (BAC-SMAD3). Following expansion, the plasmids were extracted using the Maxi/BAC’ protocol with the Nucleobond AX 100 kit (Macherey-Nagel, Hoerdy, France).
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