Figure 9

Growth inhibition after transfection with TBLR1. a. DiI staining of HK293T cells 72 hours after transfection with TBLR1 constructs. The cells were stained with DiI prior to transfection. The lipophilic dye distributes to both daughter cells after division and the loss of the dye reflects the number of times the cells have divided. The color filled curves are the computer generated model of the components of the mixed population. b. Growth of JURKAT cells over-expressing TBLR1. JURKAT cells infected with retroviral constructs expressing only GFP or GFP and a TBLR1 construct were seeded at a concentration of 25,000 per ml in 12 well Costar plates. Each day the entire contents of a well was removed and counted using a Coulter Model Zf cell counter. Each culture was performed in triplicate and the mean and S.D have been plotted. c. Cell cycle parameters (DNA content) of log phase JURKAT cells ectopically expressing TBLR1. Cells were harvested while in log phase of growth (72 hrs after plating at 50,000 cell per ml) and stained with PI as described. Fluorescence from PI bound to DNA was analyzed using a BD FACScan cytometer and the list mode data analyzed using ModFit (Verity Software) to calculate cell cycle parameters.