Figure 6. Net1 depletion reduces DEPDC7-mediated NF-κB activation.

A) Cell lysates from HEK-293 cells were immunoprecipitated (IP) with the indicated antibodies and analyzed by immunoblotting. Anti-myc antisera (Santacruz) served as a control IgG. B) HEK-293 cells were infected with retroviral vectors encoding for shRNAs targeting Net1 or a control sequence (scramble). After selection, cells were transiently co-transfected with an expression vector encoding the indicated polypeptides, together with NF-κB-luciferase and β-galactosidase reporter vectors. 24 hrs after transfection, cell lysates were prepared, and luciferase activity was measured. Data shown represent relative luciferase activity normalized against β-galactosidase activity and are representative of at least 10 independent experiments performed in triplicate. shRNAs targeting Net1 have been described in [24]. Statistical analysis was performed by Student's t test; a p value of <0.05 was considered significant.