Figure 2

Transfection efficiency of HIV-1 _YU-2 gene expression plasmid in cultured human astrocytes. To assess the transfection efficiency of cultured human astrocytes with HIV-1_YU-2, astrocytes were transfected with 0.3 μg/1.5 million cells using the Amaxa Rat Astrocyte Nucleofector kit and plated in 48 well tissue culture plates at 1₁₀ ⁵ cells/well. Three and five days post transfection cells were fixed and immunocytochemically labeled with an astrocyte marker, GFAP (green) and HIV-1p24 (red). Seven images were taken from multiple wells of HIV-1_YU-2-transfected astroctyes and the number of GFAP positive and HIV-1p24 positive cells in each image were counted independently. The number of cells positive for both markers was then calculated as a percentage. (A) Day three representative image (200x, original magnification), (B) Day five representative image (200x, original magnification) (C) Percent double positive cells following analysis of all images on both day 3 and day 5. An average 90% transfection efficient was achieved with the HIV-1_YU-2 gene expression plasmid by this method in cultured human astroctyes.