Neural Crest-Derived Dental Pulp Stem Cells Figure 12. Wnt1-marked DPSCs gave rise into neural crest-derived non-mesenchymal lineages. (A–D) Neuronal-induced cells stained positively for neuronal markers; neurofilament and S100 (in red). (E) The neurofilament staining was confirmed by the expression of neurofilament- light (NFL) and -heavy (NFH) of treated cells in neurogenic media after 21 days (N D21) (F–I) DPSCs in smooth muscle differentiation media showed smooth muscle-like phenotype that stained positively in red fluorescence for smooth muscle actin, smooth muscle myosin heavy chain, calponin, caldesmon. DAPI used for nuclei staining is depicted in blue. (J) Corresponding to immunofluorescence, Q-RT-PCR revealed the up-regulation of smooth muscle-specific genes, SRF, Sm22-alpha, Sma, SMHC, and calponin in DPSCs after cultured in smooth muscle media. Scale bars indicate PLoS ONE | www.plosone.org 13 November 2011 | Volume 6 | Issue 11 | e27526