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. 2016 Jan 19;126(2):527–542. doi: 10.1172/JCI80937

Figure 5. β2SP/SMAD3 interact with CTCF in cell nucleus.

Figure 5

(A) CTCF interacts with β2SP/SMAD3 in cell nucleus. HepG2 cells were treated with 200 pM TGF-β for 2 hours. Cell lysates were isolated as nuclear (N) and cytoplasmic (C) compartments and were immunoprecipitated (IP) with a CTCF antibody and immunoblotted (IB) with indicated antibodies. Asterisk designates nonspecific bands. (B) The interaction of CTCF and SMAD3 is TGF-β dependent. SNU398 cells were cotransfected with the indicated plasmids and were cultured with serum-free medium or treated with 5 μm SB431542 overnight. Then cells were treated with 200 pM TGF-β1 for 2 hours and cell lysates were immunoprecipitated with a CTCF antibody. Data are representative of 2 (A), and 3 (B) independent experiments. (C) Proposed model of the complex of β2SP/SMAD3/CTCF in the cell nucleus responding to TGF-β.