Additional file 1: Figure S1. of Expression of the cereblon binding protein argonaute 2 plays an important role for multiple myeloma cell growth and survival

Lenalidomide induced cell death is a slow process. Cell survival was monitored by MTT assay. Figure S2. Co-transfection of pCDH.GFP.AGO2 with pNUT.CRBN.His yielded methotrexate (MTX)-resistant cells that express high levels of GFP. Pictures were taken from MTX-selected BHK cells that were transfected with the mixed DNAs containing pCDH.GFP.AGO2 and pNUT.CRBN.His. Figure S3. AGO2 expression is associated with cell growth. Cell lysates were harvested from My5.LV without any treatment (−), treated with dimethyl sulfoxide (DMSO) or 10 μM lenalidomide (LEN) and analyzed by western blot (probed with AGO2-specific antibody). Figure S4. The growth rate of the MM cells with higher levels of CRBN is lower than the cells with low levels of CRBN. Cell survival, after infection of My5.LV or My5.CRBN.His cells with viral particles containing pCDH vector (A), AGO2 cDNA (B), pLKO.1 vector (C) or AGO2-shRNA-72 (D), was followed by MTT assay. Figure S5. Silencing of AGO2 with its shRNA or over-expression of AGO2 altered the steady-state levels of miRNAs. Total RNAs were isolated after AGO-shRNA72 (sh72) treatment for 3 days or over-expression of AGO2 and the steady-state levels of miRNAs were analyzed with microRNA array kit. Scatter plots (a), heat-maps (b) and up- or down-regulated miRNAs (c) were established by comparison of the steady-state levels of miRNAs between two samples listed in each plot. Figure S6. Treatment of MM cells with lenalidomide altered steady-state levels of miRNAs. Total RNAs were isolated after lenalidomide treatment for 3 or 5 days and the steady-state levels of miRNAs were analyzed with microRNA array kit. Scatter plots (a), heat-maps (b) and up- or down-regulated miRNAs (c) were established by comparison of the steady-state levels of miRNAs between two samples listed in each plot. DOCX 5669 kb)