Figure 6. Smad2 phosphorylation by T cell culture supernatants.

Recombinant TGF-β (1 ng/ml) or a T cell culture supernatant containing equivalent amount of TGF-β activity was added to 293T-caga-Luc-CD32-CD86 cells. After 2, 5, 15, and 30 min, the cells were lysed, and the lysates were run on SDS-PAGE. After transfer to a PVDF membrane, the membrane was blotted with anti-phospho-Smad2 Ab, and reblotted with anti-Smad2/3 Ab.