Figure 4: SM transiently affects stemness of NSC. (A) Experimental design for Rescue experiment. After 5 days of SM or 1G as control, cells were shifted to 1G for additional 5 days. Cells were also plated for clonogenic assay. (B) Percentage of secondary neurosphere-forming assay of NSC grown in SM and 1G as control, p<0.05 (C) Western blot showing Nanog levels and GAPDH as loading control in NSC grown in SM and 1G, RES: rescue. (D) Immunofluorescence staining of Nanog (green) in NSC grown in 1G, SM and after 5 days of recovery in normogravity (RESCUE). Nuclei are counterstained with hoechst (blue); scale bar 10 μm. Experiments were performed in triplicate.