SMINS-TTF-3 cells induced from mouse tail-tip fibroblasts utilizing three small molecules (Bix01294, RG108, and PD0325901). (a) SMINS-TTF-3 neurospheres were cultured in petri dishes under bright field in suspending culture. (b) A SMINS-TTF-3 neurosphere was cultured on matrigel coated dishes under bright field in attaching culture. (c-d) SMINS-TTF-3 neurospheres were dissociated and stained by typical neural stem cell markers Sox2 (red) and Nestin (red) examined by immunocytochemistry. DAPI was used for nuclei counterstaining (blue). (e) Analysis of typical neural stem cell gene expressions by RT-PCR, (1) NS (native neural stem cells), (2) SMINS-TTF-3 (small molecule-induced neural stem cells from TTF with 3 small molecules), and (3) TTF (tail-tip fibroblasts). Scale bar: 100 μm.