Fig. 4. Y-27632 potentiated CoCl₂ effect on MSC differentiation into neuron-like cells. (A,B) MSC morphological changes observed after 72 hours of Y-27632 (30 µM) and/or CoCl₂ treatment by phase-contrast microscopy (A) and fluorescence microscopy (B) after Rhodamine-labelled phalloidin staining. (C) Effect of Y-27632 with or without CoCl₂ on NSE, MAP2c and cyclin D1 expression, analysed by western blot of MSCs after 72 hours of treatment. Mouse primary-neuron-enriched cultures (11 days in vitro) were used as a positive control and actin was used as protein-loading control. (D,E) TH expression analysed by (D) immunostaining and (E) western blotting after 72 hours of Y-27632 treatment with or without CoCl₂.