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. 2013 Mar 25;3:1534. doi: 10.1038/srep01534

Figure 1. Luciferase activity in P. falciparum- infected erythrocytes transfected by electroporation (EP) and nanosome delivery.

Figure 1

(a) Optimization of K4-DNA mediated P. falciparum transfection. In [A]: K4-DNA was added to parasite pellet and resuspended in culture media. In [B]: K4-DNA was incubated with parasite pellet for 30 min at 37°C and then diluted with culture media. In [C]: K4-DNA was added to parasite pellet, resuspended in 0.5 ml incomplete medium and immediately diluted with culture media. In [D]: K4-DNA was mixed with parasite pellet resuspended in 0.5 ml incomplete media, incubated for 30 min at 37°C and then diluted with culture media. In [E]: K4-DNA was mixed with parasite pellet resuspended in 1 ml incomplete media and diluted with culture media and in [F]: K4-DNA was mixed with parasite pellet resuspended in 1 ml incomplete media, incubated for 30 min at 37°C and diluted with culture media. (b) Luciferase activity was measured 48 h after transfection for EP and four different nanosome formulations K1-K4 using plasmid pHLH-1. Formulations (K1-K4) were added to parasite pellet incubated as described in method [D] above. (c) Luciferase activity was measured 48 h after transfection for EP (50 μg) and K4 using 2.5 μg, 10 μg, 25 μg and 50 μg plasmid pHLH-1, respectively. (d) Luciferase activity was measured 48 h after transfection for EP and K4, (100% to 25% proportional reduction of K4 formulation) with plasmid pHLH-1 using method D. (e) Luciferase activity was measured after transfection by EP and K4 formulation method using 25 μg circular and linear pHLH-1. Aliquots of parasites were withdrawn every 48 hrs for luciferase activity detection and continued for up to 192 h (Day 8). (f) Luciferase activity was measured 48 h after transfection by EP and K4 formulation method using 25 μg of pHLH-1 and pLH plasmid using method D. All experiments (a–f) were performed in triplicates and vertical bars indicate standard deviation. P values were calculated using unpaired t test, and P < 0.05 indicates that there was a statistically significant difference between the formulation and electroporation luciferase activity.