Dcf 9000 gt

Transduced GSCs were seeded onto coverslips and incubated with 10 µM EdU (Toronto Research Chemicals) for 1 h at 37 °C. The cells were fixed with 4% paraformaldehyde, followed by blocking with immunofluorescence blocking buffer (10% FBS, 1% BSA, 0.3% Triton-X100). After that, the cells were stained with EdU staining solution (100 mM Tris pH 7.5, 4 mM CuSO₄, 1 mg/ml Sulfo-Cyanide Azide, 100 mM Sodium Ascorbate) for 1 h at room temperature. Images were captured with a Leica DCF 9000 GT digital camera, using a Leica DMi8 microscope. The data presented are from two independent experiments with similar results.

For immunofluorescence, GSCs, and mouse brain samples were fixed, blocked, and incubated with anti-BrdU (Biolegend, 339802), Nestin (Merck Millipore, MAB5326), or H3K27ac (Abcam, ab4729) for overnight at 4 °C. Following, secondary antibodies conjugated with Alexa 488 (Molecular Probes) or Alexa 555 (Molecular Probes) were applied. Images were captured with a Leica DCF 9000 GT digital camera, using a Leica DMi8 microscope. Data presented are from two independent experiments with similar results.