Figure 1.

Thiostrepton treatment causes downregulation of FoxM1 and its downstream targets in DLBCL cells. (A) OCI-LY19 cells were transfected with either 100 nM scrambled siRNA or 50 and 100 nM specific siRNA targeted against FoxM1 for 48 h. After incubation, cells were lysed and immunoblotted with antibodies against FoxM1, MMP-2 and MMP-9. Beta-actin was used as a loading control. (B) SUDHL4 and OCI-LY19 cells were treated with 5 and 10 μM thiostrepton for 48 h. After cell lysis, proteins were separated on SDS-Page and immunoblotted with antibodies against FoxM1, MMP-2, MMP-9 and beta-actin. (C) Clonogenic assays were performed as described in the Design and Methods section. SUDHL4 cells were treated with 5 and 10 μM thiostrepton for 48 h. Subsequently, cells were plated on Soft agar plates for four weeks. After four weeks, plates were stained and manually counted. (D) SUDHL4 and OCI-LY19 cells were treated with 5 and 10 μM thiostrepton for 48 h. Activity of MMP-2 was determined by enzyme-linked immunoabsorbent assay (ELISA). (E) SUDHL4 cells were treated with 5 and 10 μM thiostrepton for 24 h. Following treatment, Invasion-Migration assay were performed as described in the material and methods section. (F) SUDHL4 cells were transfected with either scrambled siRNA or FoxM1 specific siRNA for 48 h. Following treatment, Invasion-Migration assay were performed as described in the Design and Methods section.