FIG. 6.
Growth properties of ROD1 overexpressors. (A) Level of Rod1 expressed in the stable ROD1 transfectants. The human ROD1 cDNA inserted into the CMV promoter-based expression vector (27) was transfected into K562 cells by electroporation. Stable transfectants were selected in medium containing Geneticin (G418 sulfate) at 1.5 mg/ml as described in Materials and Methods. Cell lysates were prepared from each stable transfectant, and Rod1 protein was detected by Western blotting with the anti-human Rod1 monoclonal antibody 2B3. Rod1-C2, Rod1-C3, Rod1-C7, and Rod1-C9 are transfectant clones expressing high levels of Rod1. Neo1 to Neo10 are also stable transfectants of an empty vector and are used as a negative control. (B) ROD1 overexpression does not affect the growth rate. Cells were plated at 105 cells/ml and incubated to monitor the growth of each Rod1 overexpressor. The cell number was counted at the indicated times. The values in the figure are the means ± the standard deviation for the Rod1 overexpressors and for Neo1, -2, -5, and -7.