(a)
FOXM1 message levels in H929 and ARP1 myeloma cells, using qRT-PCR as measurement tool. Cells either under-expressed FOXM1 due to lentiviral transduction of a FOXM1–targeted shRNA “knockdown” construct (KD) or expressed FOXM1 at normal levels (N) following transduction of a non-targeted or “scrambled” shRNA used as control. Average loss of FOXM1 mRNA upon gene KD was ~80% and ~70% in H929 and ARP1 cells, respectively.
(b) Western analysis of samples included in panel a. Whole cell lysates were electrophoretically fractionated and immunoblotted using antibodies to FOXM1 and β-actin. Densitometry was employed to determine the FOXM1-to-β-actin ratio. Loss of FOXM1 protein in H929 and ARP1 KD cells amounted to 74% and 59%, respectively.
(c) Photographic images of representative soft-agar plates indicating the decreased clonogenic growth of FOXM1KD cells (bottom) compared to FOXM1N cells (top).