Figure 5.

Spiropyrazoline oxindole 1a decreases NSC self-renewal through a cell cycle independent manner. Mouse NSCs were treated with compound 1a at 12.5 μM or DMSO (control) for 24 h and collected for flow cytometry and cell-pair assays as described in Experimental. (A) Effect of compound 1a on cell cycle progression. Cellular DNA was stained with propidium iodide (PI) in differentiation conditions to determine cell cycle distribution. (B) Representative quantification data of paired cells Sox2-positive and -negative, in low EGF/bFGF containing medium after 1a treatment. (C) Representative microscopy images of undifferentiated NSC pairs labeled with Hoechst 33258 for nuclei staining and anti-Sox2 antibody after 1a treatment. Scale bar, 25 μm. Results are expressed as mean ± SEM fold-change for at least three different experiments. ^*p < 0.05 from control by Student's t-test.