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. 2012 Feb 3;1(3):247–260. doi: 10.1242/bio.2012463

Fig. 7. Endogenous Smad7 regulating an E2F-responsive promoter.

Fig. 7.

(A) Association of endogenous Smad7 and HDAC-1 proteins in A549 human lung cells. Nuclear extract from untransfected A549 cells were incubated with an α-HDAC-1 antibody or control IgG. Immune complexes were examined by Western blotting using α-Smad7 and α-HDAC-1 antibodies. (B) Association of endogenous Smad7 and E2F-1 proteins. Immune complexes prepared using α-E2F-1 or control IgG as in (A) were examined by Western blotting using α-Smad7 and α-E2F-1 antibodies. (C) Effect of Smad7 knock-down on the transcriptional activation of E2F-responsive promoter. (Upper panel) In combination with a vector for either Smad7-targeting or control siRNA, luciferase reporter (pGVB2-E2F×4) driven by an E2F-responsive promoter was transfected into A549 cells. Cells were serum-starved for 40 h, stimulated in medium containing 10% FBS for 18 h and then lysed for assays. Luciferase activity in the lysates is shown as means with S.D. from triplicate experiments. (Lower panel) Level of endogenous Smad7 in the lysates was examined by Western blotting using α-Smad7.