FOXM1 and interactions with the MMB complex. (A) GST pulldown analysis using GST or GST-FOXM1(1–367) and the indicated in vitro translated MMB and DREAM complex components. Precipitated and input in vitro translated proteins were detected by phosphorimaging (top panel) and GST fusion proteins were detected by Coomassie blue staining (bottom panels). Arrow represents the position of the band corresponding to full-length GST-FOXM1(1–367). (B) GST pulldown analysis using GST or GST-FOXM1(1–367) and the indicated bacterially expressed and purified full-length and deleted Flag-tagged Lin9 proteins (shown diagrammatically at the top). FOXM1 and Lin9 derivatives were detected by immunoblotting with anti-Flag (top) or anti-GST antibodies (bottom). The region of Lin9 which is sufficient for FOXM1 binding is boxed. (C) Nanostring nCounter gene expression assay of the indicated control or FOXM1 target genes following siRNA-mediated knockdown of FOXM1 or LIN9. Data are shown as a heat map of fold change (log2) relative to a non-targeting siRNA control and are the average of three independent experiments. Grey dots represent the identities of genes in the overlap of the Venn diagram in part (D). (D) Venn diagram of genes showing significant (p < 0.05, t-test) changes in expression following knockdown of one of the two indicated factors. Note that ARHGAP19 changes in opposite directions for each knockdown.