Fig. 2.

The transfection efficiency of human astrocytes. Primary human astrocytes were transfected using nucleofection technology. CD38 siRNAs were labeled with Cy3, and labeled nucleic acids were precipitated to remove free Cy3. Transfected cells were cultured for 12 h and activated with IL-1β (20 ng/ml) for 10 h (B, total 22 h post-transfection) and 24 h (D, total 36 h post-transfection). Parallel controls without IL-1β-activation were also evaluated (A, C, and E). Ten independent fields were photographed with 90–100 cells per field and Cy3-positive cells versus total cells were counted. Subpanel F shows the cumulative percent of Cy3-positive cells in each condition. On an average, 85% of the cells were transfected at all time points and IL-1β activation had no effect on transfection efficiency. Original magnification (×200)