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. Author manuscript; available in PMC: 2010 Nov 1.
Published in final edited form as: Mol Carcinog. 2009 Nov;48(11):1048–1058. doi: 10.1002/mc.20557

Figure 5.

Figure 5

Ser145 Phosphorylation of MAD1 inhibits its transcription repression to target genes. (A) MAD1(S145A) mutant transcription repression was not affected by AKT. Lysates of MDA-MB-435 cells transfected with Flag-MAD1 or Flag-MAD1(S145A) mutant with or without AKT were subjected to reporter assay. (B) mRNA levels of MAD1 target genes hTERT and ODC were affected by the presence of AKT in MAD1 but not in MAD1(S145A) mutant. RNA of MCF7 cells transfected with the indicated plasmids were subjected to quantitative PCR. (C) MAD1(S145A) mutant did not reduce its suppression of the cell cycle in the presence of AKT. MCF7 cells were transfected with the indicated plasmids, and GFP-positive cells were gated for cell cycle assay by flow cytometry. (D) The repression of cell growth by MAD1(S145A) mutant was not affected by AKT. Hela cells were transfected with wild-type MAD1 (WT) or mutant MAD1 (S145A) in the present or absence of AKT. The growth rate was measure by MTT assay.