Flow Cytometry Gating Strategy Used to Identify EdU+ and p-Ser10-H3+ Glioma Cells
(A) Total cells are gated to exclude cellular debris (top left plot). Then, doublet discrimination gating is performed to filter out cellular aggregates (top middle and right plots). Lower panel: Left histogram: glioma cells positive for EdU (black=unstained control, blue=EdU (Pacific Blue staining)). Right histogram: glioma cells positive for p-Ser10-H3 expression (black=unstained control, red=p-Ser10-H3 (PE staining)).
(B) Representative results for p-Ser10-H3 and EdU staining of tumor cells isolated from SB gliomas. Tumors were generated by the introduction of the following genetic alterations: NRASG12V overexpression, p53 and Atrx silencing for NPA; or NRASG12V overexpression, p53 and Atrx silencing, and IDH1R132H overexpression for NPAI (Table 1). Results are shown as the % of positive cells (left table) or the median fluorescent intensity (MFI) (right table) ± SD, for each staining. NPAI tumors are less aggressive than NPA tumors and cells show different cell cycle characteristics.