American Society of Hematology

Fig. 1.

$Fig. 1. Flow cytometric analysis of transgene expression in K562 cells following EGFP mRNA electroporation. / (A) K562 cells were electroporated with EGFP mRNA at 300 V, 150 μF or with EGFP plasmid DNA at 260V, 1050 μF (dashed line) as described in “Materials and methods.” Twenty-four hours after electroporation, flow cytometric EGFP analysis was performed to estimate transfection efficiency of mRNA electroporation (bold line) and plasmid DNA electroporation (dashed line). An overlay histogram representative of 5 independent experiments is shown. Nonelectroporated cells (thin line) were used to determine background fluorescence. The M1 region indicates the EGFP-positive cell fraction. The percentage of EGFP+cells was 85% (bold line) and 50% (dashed line) following mRNA or plasmid DNA electroporation, respectively. (B) Kinetics of EGFP mRNA expression in K562 cells in function of time (n = 3). Note the rapid induction of high-level EGFP expression already 3 hours following electroporation.$

Flow cytometric analysis of transgene expression in K562 cells following EGFP mRNA electroporation.

(A) K562 cells were electroporated with EGFP mRNA at 300 V, 150 μF or with EGFP plasmid DNA at 260V, 1050 μF (dashed line) as described in “Materials and methods.” Twenty-four hours after electroporation, flow cytometric EGFP analysis was performed to estimate transfection efficiency of mRNA electroporation (bold line) and plasmid DNA electroporation (dashed line). An overlay histogram representative of 5 independent experiments is shown. Nonelectroporated cells (thin line) were used to determine background fluorescence. The M1 region indicates the EGFP-positive cell fraction. The percentage of EGFP⁺cells was 85% (bold line) and 50% (dashed line) following mRNA or plasmid DNA electroporation, respectively. (B) Kinetics of EGFP mRNA expression in K562 cells in function of time (n = 3). Note the rapid induction of high-level EGFP expression already 3 hours following electroporation.

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