Figure 1.

Schematic representation of the experimental procedure. Animals were habituated for 2-3 days in the CaResS device before sleep deprivation. After 6 h of sleep deprivation, mice were sacrificed by decapitation and cerebral cortices were dissociated. Cell suspension was purified by FACS in 2 populations: GFP-positive/PI-negative cells and GFP-negative/PI-negative cells, based on sequential gating of size, viability (PI fluorescence or y-axis on the graph), and GFAP-GFP fluorescence (x-axis on the graph), where small dots represent sorted events (≈cells). RNA samples from 3 animals were pooled to obtain sufficient RNA for one replicate, and 5 replicates per experimental group were used to quantify mRNA levels. FACS, fluorescence activated cell sorting; GFP+ cells, green fluorescent protein expressing cells; GFP- cells, cells that do not express green fluorescent protein; PI, propidium iodide.