TGFβ-induced suppression of CD248 is mediated via canonical signaling pathways. (A, B, C) MEF were incubated for 48 hrs with TGFβ 3 ng/ml and the ALK-inhibitor SB431542 1 μM either singly or in combination. Controls included carriers for SB431542 (DMSO) or for TGFβ (0.1% BSA). (A) Western blots and (B) immunofluorescence were used to detect expression of CD248 (green). (C) CD248 mRNA levels were also quantified (n = 3 experiments, each in triplicate; *p < 0.05). Results indicate that TGFβ-mediated suppression of CD248 protein and mRNA requires integrity of canonical ALK5-Smad2 signaling pathway. Scale bar = 50 μm.