Figure 5

Effect of Hedgehog blockade upon neurosphere formation. The ability to form neuropheres in the presence of the Hedgehog inhibitor cyclopamine was evaluated in Daoy cells. Spheres derived from Daoy control (upper) and Msi1 knockdown (KD; lower) were incubated for one week in the presence of inhibitor (Cyc) and tomatidine (Tom; negative control) at a concentration range of 5–20 μM. A reduction in neurosphere size was observed in both cell lines; however, the knockdown cell line appeared to be more sensitive to cyclopamine treatment as the effects upon sphere formation were significant (p < 0.05) at lower concentrations of inhibitor. a) Representative images of the effects upon neurosphere formation at 5 μM and 20 μM cyclopamine are shown. Scale bars represent 100 μm. b) The average number of neurospheres per field is represented for control and KD cells after treatment with tomatidine and cyclopamine. * indicates p < 0.05, ** p < 0.01 and *** p < 0.001.