fig1

Figure 1. Glutaminolysis in U266 cells and GLS1 and GLS2 expression in monoclonal gammopathy of undetermined significance and MM cells. (A) The viability of cells was evaluated by culturing them in medium with or without Gln for 72 h; (B) Caspase 3/7 activity was measured; and (C) intracellular ATP levels were quantified. Statistical significance is denoted as ^***P < 0.001 and ^****P < 0.0001 compared to the complete medium cultured cells. ns: not significant; (D) U266 cells were cultured in RPMI 1640 medium, without Gln or without Gln plus glucose, for 72 h. Cell viability was analyzed. ns: not significant; (E and F) GLS1 and GLS2 expression levels were validated using data from the GEO database. Statistical significance compared to normal is indicated. ns: not significant. (G and H) Total RNA was extracted from normal PBMC and myeloma cell lines. GLS1 and GLS2 gene expressions were analyzed using RT-PCR. GLS: Glutaminase; MM: multiple myeloma; ATP: adenosine triphosphate; RPMI 1640: Roswell Park Memorial Institute 1640; GEO: Gene Expression Omnibus; PBMC: peripheral blood mononuclear cell; RT-PCR: reverse transcription-polymerase chain reaction.