Fig. 2.

Single GBM NG2+ and GBM NG2− cells generate progeny that establish phenotypic equilibrium. (A) Experimental design for (B) and (C). Single NG2+ (red) or NG2− cells (blue) are seeded in 96-well plates, then tested for clonogenic capacity and NG2 expression by their progeny. (B) NG2+ cells demonstrate increased clonogenicity at all timepoints. Micrographs show examples of colonies formed from single NG2+ or NG2− cells. (C) Progeny of single GBM NG2+ and GBM NG2− cells generate mixed populations of NG2+ and NG2− cells. Illustrative example from G25 cell line. (D) Design of the experiment in (E). Unsorted GBM cells (W-GBM, red and blue) are stably transfected with GFP to form G-GBM population (green center). Single NG2+ (red) and NG2− (blue) cells from the G-GBM population are sorted into plates containing W-GBM cells. (E) Micrographs and flow cytometry analyses of the progeny of: (left) G-GBM NG2+ mixed with W-GBM cells and (right) G-GBM NG2− cells mixed with W-GBM cells. In both cases, the sorted G-GBM cells produce NG2+ and NG2− cells.