Canonical Wnt Pathway Inhibitor ICG-001 Induces Cytotoxicity of Multiple Myeloma Cells in Wnt-Independent Manner
Fig 4
Involvement of Wnt signaling in ICG-001-induced apoptosis in MM cells.
(A, B) MM MM.1S cells were transfected with siRNA against β-catenin or control non-targeting pool siRNA. (A) The efficiency of knockdown was evaluated by Western blotting with antibody against β-catenin. Equal loading was confirmed by re-probing the membrane with antibody against β-actin. (B) Cells were then either left untreated (control) or were treated with indicated concentration of doxorubicin for 24 hours. Apoptosis of MM cells was detected by Annexin V/DAPI staining by flow cytometry. Three experiments were performed with similar results. Results of 1 representative experiment are shown. (C, D) MM.1S cells were transfected with pcDNA/Myc TCF4 or control empty pcDNA3 plasmid followed by selection of stably transfected clones with G418. (C) Overexpression of TCF4 was evaluated by Western blotting using anti-Myc-Tag antibody. Equal loading was confirmed by re-probing the membrane with antibody against β-actin. (D) MM.1S cells transfected with pcDNA/Myc TCF4 or empty pcDNA3 vectors were treated with ICG-001 (10 μM) for 24 hours followed by detection of apoptosis by Annexin V binding assay. Combined results of 2 independent experiments are shown.