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. 2012 Dec 7;7(12):e50815. doi: 10.1371/journal.pone.0050815

Figure 4. ΔNp73 and Smads form a complex with SBEs in vitro.

Figure 4

A) DNA Affinity Precipitation assay (DNAP) of SBE DNA duplexes with extracts of cells left untreated or treated with TGF-β1 after triple transfection with YFP-tagged Smad4, V5-tagged Smad3 and either empty vector control (c), HA-tagged ΔNp73 (ΔN) or HA-tagged TAp73α (TA). B) Relative binding of V5Smad3 to SBE oligos, quantification and normalization for input values of the V5Smad3 results from Figure 3D. C) Relative binding of YFPSmad4 binding to SBE oligos, quantification and normalization for input values of the YFPSmad4 results from Figure 3D. D) Pull-down of SBE oligo incubated with a cell lysate containing HA-ΔNp73 (extract 1) or with a cell-lysate containing V5Smad3 and YFPSmad4 (extract 2) or incubated first with extract 1, washed and incubated with extract 2 (1+2). E) Immunoprecipitation (IP) with α-HA of HA-ΔNp73 induced cells (tetracycline) detecting endogenous Smad3.