Figure 4

Efficiency of cap-dependent versus internal ribosome entry site-dependent translation in various cell lines. Plasmids pEF3-ChFP (in red), pEF3-EGFP (in green), pEF3-EGFPEMCVChFP (in black) and pEF3-ChFPEMCVEGFP (in brown) were transfected into (a) 293T cells, (b) B16 cells and (c) mesenchymal stem cells (MSCs). Contour plots of FL1 (EGFP levels) versus FL3 (monomeric cherry fluorescent protein (ChFP) levels) were made of the green and red fluorescence from a sample of cells in each population. Lines were drawn through the medians of the cells demonstrating strong EGFP and ChFP fluorescence. (d) Because insufficient numbers of cells with strong fluorescence could be obtained in MSCs transfected with these plasmids, plasmids pmaxCDK-EGFPEMCVChFP (black) and pmaxCDK-ChFPEMCVEGFP (brown) were transfected into MSCs. Each population was translated in the dotplot so that the populations of nonfluorescent cells optimally overlapped.