Figure 8.

Setup of transfections of MGE-derived cells. (A,B) Test of different conditions of MGE–derived cells plated on 200 μg/ml PLL and 20 μg/ml LN, transfected at DIV1, and fixed at either DIV4 or DIV6 for immunofluorescence. After transfection cells were cultured without (A) or with 50 ng/ml BDNF (B). (C) Quantification of the percentage of cells transfected at DIV1 with 0.1 μg of the indicated plasmid DNA and 0.5 μl of Lipofectamine-200, and fixed at different DIVs for immunofluorescence. Bars are means ±SEM of percentage of DAPI-positive cells expressing GFP. (D) Immunofluorescence of a culture transfected with GFP grown in the presence of BDNF, stained with Abs for GFP (green), for MAP2 (red), and with DAPI (blue). (E) DIV6 cells cotrasfected at DIV1 with 0.05 μg of each plasmid (total of 0.1 μg of plasmid DNA), to express GFP and HA-tagged guanine nucleotide exchange factor PIX.