Chunk #52 — MATERIALS AND METHODS — Fluorescence-activated cell sorting

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Musculocontractural Ehlers-Danlos syndrome and neurocristopathies: dermatan sulfate is required for Xenopus neural crest cells to migrate and adhere to fibronectin.

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To assess cell viability, dissociated cells from 20 CNC explants were incubated with 10 μg of propidium iodide (PI) and analyzed ∼2 h after extraction by performing FACS, with BD FACSCalibur™ and BD cellQuest™ pro software for acquisition. Analyses were performed with Flojow version X. To monitor the cell cycle, dissociated CNC cells were fixed with ethanol at 4°C overnight. The cells were subsequently washed and incubated with 10 μg of RNase A for 15 min. Prior to the analyses, the cells were incubated with 10 μg PI.