(A) Soft agar colony-forming assay to determine clonogenicity of SSEA-1+ versus SSEA-1− cells from three different GBM lines.
(B) Limiting dilution sphere-forming assay. Three cell lines used in (A) were plated into 96-well plates with various seeding densities (5–50 cells per well, 30 wells per each condition). Data from a representative experiment were shown. R2 > 0.83.
(C and D) PCNA staining (C) and cell-cycle analysis (D) of 1228 and 0131 cells by flow cytometry. Error bars represent SD (performed in triplicates); *p < 0.05.