Direct reprogramming of C6 glioma to neurons using two small molecules.

(A) Representative fluorescence images of small molecule-induced neurons (SMiNs) 35 days after neural induction with CHIR99021 (20 μM) and forskolin (100 μM). (B) Quantitative result of Tuj1, MAP2, GFAP, PDGFR, and NG2 staining in SMiNs 35 days after neural induction. (C) Representative image of Tuj1- or MAP2-positive SMiNs 35 days after neural induction with CHIR99021 (20 μM) and forskolin (100 μM). (D) Quantitative result of morphological complexity of Tuj1-positive SMiNs 35 days after neural induction. (E) Representative band image of SSEA1 (a cancer stem cell marker) and C-Myc (an oncogene marker) 35 days after neural induction with CHIR99021 (20 μM) and forskolin (100 μM). (F) Representative fluorescence images of SMiNs 7 days after neural induction with lithium carbonate (3 mM) and forskolin (100 μM). (G) Representative fluorescence images of SMiNs 7 days after neural induction with CHIR99021 (2 μM) and dbcAMP (0.5 mM). (H) Representative images of patch-clamp in SMiNs. (I) Representative traces of sodium current of SMiN 84 days after neural induction before (left) and after (right) 0.5μM TTX bath application. (J) Representative traces of action potential spike of SMiN 84 days after neural induction before (left) and after (right) 0.5μM TTX bath application. (K) Representative traces of sodium current (left) and action potential spike (right) of undifferentiated C6 glioma in proliferation media. (L) Representative traces of sodium current (left) and action potential spike (right) of undifferentiated C6 glioma in neural differentiation media without small molecules. Data are presented as the mean ± S.E.M. Scale bars represent 50 μm (A), 20 μm (C), 100 μm (F), and 100 μm (G).