Expression of CD248 in response to increasing concentrations of TGFβ. (A) MEF were incubated for 48 hrs with increasing concentrations of TGFβ. Expression of CD248 (seen as monomers (~160 kDa) and dimers) and phosphorylation of Smad2, were detected by Western blot. (B) CD248 expression relative to actin expression was quantified by densitometry (n = 3 experiments) and results were normalized to the no-treatment condition. (C) CD248 expression by MEF (wild-type, WT; or lacking CD248, KO) was detected with specific anti-CD248 antibodies after exposure to carrier (Control) or TGFβ for 48 hrs. TGFβ suppresses CD248 in a concentration-dependent manner, with simultaneous increase in phosphorylated Smad2 and ERK1/2. Scale bar = 50 μm.