Figure 2.

Addition of DOPE/CHEMS significantly enhanced transfection. (A) Polyplex of LPEI/pDNA (N/P = 10, 2 μg of pDNA) was added to differentiated Neuro2A cells (10 μM RA) and centrifuged at 280 g (5 min). After which, transfection mixture was replaced with fresh media. One hour post-transfection, 50 μl of 25mM HEPES (–) or 12 μg of DOPE/CHEMS prepared at 9:2 molar ratio in 25 mM HEPES (+) was added to the culture medium. Twenty-four hours later, transfection efficiency was analyzed by counting the fluorescent and bright field images. To determine the transfection efficiency of the differentiated population, cells bearing neurites twice the cell body length were counted. Transfection efficiency was calculated as the percentage of EGFP positive cells normalized to the total number of differentiated cells per image. Data are shown as mean ± SEM of biological triplicates. Significant differences between data points were calculated using two tailed Student's t-test. **P < 0.005. (B) Representative images captured (20x magnification) at the end of incubation were presented.