ChIP was performed as described (40). Briefly, cells were cross-linked with 1% formaldehyde for 20 min before being sonicated using output power 3 (9-W power) for 10 cycles. Before the immunoprecipitation, Dynabeads Protein G (Invitrogen) magnetic beads were washed and coupled to 10 μg of anti-SMAD4 (sc-7966, Santa Cruz Biotechnology), and normal mouse immunoglobulin G (IgG) (sc-2025, Santa Cruz Biotechnology). Cell lysates were incubated with SMAD4 and IgG-conjugated magnetic beads for overnight at 4°C. Immunoprecipitated lysates were treated with proteinase K and incubated for overnight at 65°C before the DNA being extracted.

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