Fig. 4

Wnt3A and Rspo2 maintains stemness in GSCs. a Rspo2 and Wnt3A upregulates stem cell marker in U251 GSCs as demonstrated by real time PCR. Blk indicates U251 GSCs cultured in GSC media with 0.1% DMSO. b All-trans retinoic acid (10 µM RA) was used to induce differentiation in U251 GSCs for 24 h with or without Wnt ligands (20 ng/ml). Real-time PCR was used to determine the effect on differentiation. Results show that Rspo2/Wnt3A treatment rescues RA-induced GSC differentiation. Blk indicates U251 GSCs cultured in DMEM with 0.1% DMSO. c GSCs were cultured in GSC media, or GSC media without EGF and FGF, or GSC media without EGF and FGF but with Wnt3A and Rspo2 for 7 days. Results show that lack of EGF/FGF causes reduced sphere formation, which can be compensated by adding Rspo2/Wnt3A. d Real-time PCR shows that Rspo2/Wnt3A treatment abolishes the downregulation of stem cell markers and upregulation of differentiation markers caused by growth factor deprivation. Blk in C and D indicates U251 GSCs cultured in GSC media with 0.1% DMSO