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Fig. 3. Assays for TGFß responsiveness in vitro. (A,B) Schematics of in vitro assays. (A) Keratinocyte monolayers and stratified cultures used in panels C-M; PC, postconfluence. (B) Monolayers combined with keratinocytes that had been induced to differentiate in suspension. (C-J) Immunolocalization of Smad2/3. (K,L) Immunolabelling for {alpha}6 transgene. Bar, 50 µm. (M) Western blots of untreated (-) or TGFß1-treated (+) cells probed with antibodies to total Smad2/3, activated Smad2 (pSmad2), TGFßRI or, as a loading control, actin; M, monolayer, S, stratified cultures.