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. 2016 Jun 7;5(6):e322. doi: 10.1038/mtna.2016.34

Figure 3.

Figure 3

Cell survival, transfection, cell death mechanism and morphology of B16.F10 cells after pDNA electrotransfer. Commercially prepared vector plasmid (gWiz Blank) was electrotransferred into B16.F10 melanoma cells. Control, 40 µl of cells and 10 µl of saline; pDNA 1 mg/ml, 40 µl of cells and 10 µl of 1 mg/ml gWiz Blank plasmid; pDNA 2 mg/ml, 40 µl of cells and 10 µl of 2 mg/ml gWiz Blank plasmid; pDNA 3.5 mg/ml, 40 µl of cells and 10 µl of 3.5 mg/ml gWiz Blank plasmid; EP, 40 µl of cells and 10 µl of saline were electroporated by the delivery of eight 5 ms pulses with a voltage-to-distance ratio of 600 V/cm with a plate electrode; EP+pDNA 1 mg/ml, 40 µl of cells and 10 µl of 1 mg/ml gWiz Blank plasmid were electroporated as described in EP group; EP+pDNA 2 mg/ml, 40 µl of cells and 10 µl of 2 mg/ml gWiz Blank plasmid were electroporated as described in EP group; EP+pDNA 3.5 mg/ml, 40 µl of cells and 10 µl of 3.5 mg/ml gWiz Blank plasmid were electroporated as described in EP group; EP1, 40 µl of cells and 10 µl of saline were electroporated by the delivery of six 100 µs pulses with a voltage-to-distance ratio of 1300 V/cm and frequency 4 Hz with a plate electrode; EP1+pDNA 1 mg/ml, 40 µl of cells and 10 µl of 1 mg/ml gWiz Blank plasmid were electroporated as described in EP1 group; EP1+pDNA 2 mg/ml, 40 µl of cells and 10 µl of 2 mg/ml gWiz Blank plasmid were electroporated as described in EP1 group; EP1+ pDNA 3.5 mg/ml, 40 µl of cells and 10 µl of 3.5 mg/ml gWiz Blank plasmid were electroporated as described in EP1 group. For image B, 40 µl of cells and 10 µl of 2 mg/ml pEGFP-N1 plasmid were electroporated as previously described. (a) Cell survival was measured 72 hours after electrotransfer and was normalized to the control group. (b) Transfection efficiency of B16.F10 cells after pEGFP-N1 electrotransfer after pDNA delivery with EP and EP1. *P < 0.05, statistically significant difference compared to all control groups and EP only. a.u., arbitrary units. (c) Cell death mechanism as quantified by flow cytometery and (d) cell morphology after pDNA electrotransfer. Blue arrows indicate necrotic cells displaying fragments of cytoplasm and only an outline of the nucleus. Black arrow indicates apoptotic cells with vacuolization of the cytoplasm and formation of apoptotic bodies.