mouse bmsc line  (iCell Bioscience Inc)

Journal: PLoS ONE

Article Title: GLIPR1 expression is reduced in multiple myeloma but is not a tumour suppressor in mice

doi: 10.1371/journal.pone.0228408

Figure Lengend Snippet: ( A ) The expression of Glipr1 mRNA was assessed by RT-PCR in CD138 + PCs from healthy WT (C57BL/6) mice and KaLwRij mice, as well as the KaLwRij-derived 5TGM1 MM PC line. The products were run on a 2% agarose gel and stained with GelRed. Actb was used as a positive control. NTC = no template control. ( B ) The expression of Glipr1 protein in 5TGM1-Glipr1 (Glipr1) and control 5TGM1-EV (EV) cells was assessed by Western blot. Hsp90 was used as the loading control. The number of 5TGM1-Glipr1 cells in mono-culture ( C ) or co-culture with OP9 bone marrow stromal cells ( D ) was assessed by measuring luciferase activity after 3 days. Cell number is expressed relative to the EV control cells. ( E ) Colony formation by 5TGM1-Glipr1 cells versus 5TGM1-EV cells was assessed in semi-solid methylcellulose-containing medium after 12 days. Colony number is expressed relative to the EV control cells. Graphs depict the mean + SD of n = 3 independent experiments.

Article Snippet: The mouse BM stromal cell (BMSC) line OP9 was obtained from the ATCC and was maintained in Dulbecco's Modified Eagle Medium (DMEM) with 10% FCS.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Agarose Gel Electrophoresis, Staining, Positive Control, Control, Western Blot, Co-Culture Assay, Luciferase, Activity Assay