For FACS analysis live astrocytes were isolated at room temperature by FACS at the Stanford Shared FACS Facility on the basis of their GFP expression on a BD Aria II or BD Influx. Cell suspensions were sorted twice sequentially using forward light scatter and SSC to gate single cells, followed by gating for GFP fluorescence in the absence of LIVE/DEAD stain to select live astrocytes. Individual cells were collected directly into 96-well PCR plates containing RT-STA Mix solution (see below) for microfluidic qPCR. Flowjo software (Treestar) was used to analyze purity of final astrocyte populations.
