Fig. 5

UBE2O is downregulated in MM cells and suppresses c-Maf transcriptional activity. a Relative UBE2O expression in the process of MM development. The mRNA expression levels of UBE2O were retrieved from a DNA microarray with 99 primary bone marrow samples including normal donors, MGUS, SMM, and MM patients as described in the “Methods” section. b The UBE2O mRNA levels from primary samples were analyzed by RT-PCR and agarose gel electrosphoresis. c Quantitative analyses of UBE2O mRNA from healthy donors and MM patients from (b). d UBE2O was co-transfected into HEK293T cells with a c-Maf plasmid and a luciferase reporter driven by a CCND2 promoter containing c-Maf recognition element (CCND2.Luc). e Lentiviral UBE2O was infected into MM cell lines RPMI-8226 and LP1 for 72 h. The cell lysates were then prepared for immunoblotting assay against specific antibodies as indicated. f RPMI-8226 and LP1 cells were infected with lentiviral UBE2O for 48 h followed by PI staining and cell cycle analysis