Figure 2
From: Novel strong tissue specific promoter for gene expression in human germ cells
Effect of mNUS-RCD cassette on cell viability. Cells were transfected with no-RCD (negative control), mNUS-RCD (analyte) and CMV-RCD (positive control) constructs and grown on 5FC-containing medium for 24 hours. Transfection efficiencies were measured using cotransfection with GFP-encoding vector TurboGFP (Evrogen), only transfections with efficiency values 65-80% were scored. (A), representative Western blot results with the antibodies specific to RCD and to a housekeeping protein GAPDH. (B), representative results of the propidium iodide (PI) assay. Histograms for Tera-1 cells are shown in red, for HEK 293 - in blue. Proportions of dead cells are given in percents. (C), representative FACS data for the Annexin V assay. Incidences of the living Tera-1 cells are shown in lower left quadrant (proportion in percents - bottom, left), of apoptotic cells - in lower right quadrant (proportion - bottom, right), of necrotic cells - in upper right quadrant (proportion - top, right). Overall proportions of dead cells were calculated as the total of proportions of apoptotic and necrotic cells.