Fig. 5

Silencing of AGO2 induced cytotoxicity in IMiD-resistant and IMiD-sensitive MM cells. a & b The effects of silencing AGO2 with AGO2-shRNA on MM cell growth and survival. IMiD-resistant My5.LV cells (a) and IMiD-sensitive My5.CRBN cells (b) were infected with lentiviral particles harboring either lentiviral vector, AGO2-shRNA or AGO2 cDNA. The media were replaced 18 hours post-infection and then 1 μM puromycin (final concentration) was added 24 hours later (day0). Cell survival was followed by MTT assay. c & d The effects of silencing AGO2 with AGO2-shRNA on the steady-state levels of AGO2, CRBN and β-actin. IMiD-resistant My5.LV cells (c) and IMiD-sensitive My5.CRBN cells (d) were lysed three days post infection with viral particles indicated in (a) and (b) and 100 μg of total proteins from these treated cells were loaded in each lane and analyzed by western blot. e The effects of silencing AGO2 with AGO2-shRNA on other MM cell lines. f Expression of recombinant AGO2 can reverse the AGO2.shRNA induced cytotoxicity. The infection of My5.LV and My5.CRBN cells with viral particles harboring AGO2-shRNA-72 was performed exactly the same as in (a) and (b) The rescue experiments were performed by: 1) collecting the MM cells treated with AGO2-shRNA-72 for six days; 2) infecting these cells with viral particles harboring AGO2 cDNA; 3) replacing the media 18 hours post-infection; 4) performing MTT assay 24 hours later (day1)