PI staining for flow cytometry was performed according to the user manual of DNA Content Quantitation Assay (Cell Cycle) from Solarbio (#CA1510). Briefly, OPCs from control or Tet1 cKO mice were harvested, washed in PBS, and fixed in cold 70% ethanol for 30 min at 4 °C. After washing twice in PBS, cells were treated with RNase and then stained with PI. With guava easyCyte6HT (Millipore), the forward scatter (FS) and side scatter (SS) were measured to identify single cells. For analysis, ModFit LT 5.0 software was used to make the PI histogram plot. Experiments were replicated three times.
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