Fig. 3: Tpl2^−/− primary keratinocytes are refractory to TGFβ mediated growth inhibition despite having normal levels of TGFβ receptors and SMAD proteins.

Primary keratinocytes were cultured in low calcium (0.05 mM) growth media or transduced with v-ras^Ha (RAS). Real-time PCR (a) and Western analyses (b) were conducted for TGFB, TGFBRI and TGFBRII, SMAD2 and SMAD4. c Fibroblast protein was collected from newborn WT and Tpl2^−/− mice and used for Western analysis. All expression was normalized to a housekeeping gene. d WT and Tpl2^−/− keratinocytes were treated with 10 ng/ml TGFβ and after 24 h, proliferation was determined using a WST-1 assay. e Whole mRNA was isolated and real-time PCR performed and analyzed for relative abundance of Fas mRNA