(A and B), Consistent survival of U87 (A) or U251 (B) -derived GSC clones in the methylcellulose-containing growth media throughout the culture period. The numbers of survived clones are shown at each time point (0, 4, 7 days for A and B; 14, 21, 28 days for B, respectively). The data for 5 generations are shown (1st shown in black filled circles; 2nd: blue; 3rd: green; 4th: orange; 5th: red, respectively). Clones remained to survive consistently during the assayed periods at every generation. C-N, Dose dependent suppression in the survival of clones in the presence of OTS964. The numbers of clones in the population of all clones (C–E); of self-renewed clones (F–H for clones with multiple cells); of expanded clones (I–K for clones with more than 4 cells; L–N for clones with more than 9 cells) are shown at various concentrations of OTS964 (0, 20, 100, 200, 300, 500 nM). The graphs are for U87 (C, F, I and L) and for U251 (D, E, G, H, J, K, M and N) -derived GS clones. The data of 20, 100 and 500 nM were taken from the first experiment; of 200 nM, the second; of 300 nM, the third and the fourth and the control (0 nM) from both of the first thorough the fourth. OTS964 suppressed the survival of GS clones, and the effects were dose-dependent.