Figure 6

Expression of midbrain markers by Sox1 -GFP⁺ neural progenitors and their differentiated progeny. Sox1-GFP^pos neural progenitors, generated by co-culture with PA6 (A-C, D, E, G, H, J, K) or from monolayer culture (F, I, L-O), were purified by FACS. Cells were either collected onto slides by cytospin (A-C) or re-plated on PA6 or PDL/laminin and allowed for differentiation for a further 7 days (D-O). Cells collected on slides were examined for the expression of midbrain dopaminergic progenitor markers by immunostaining using antibodies against FoxA2 (A), En1 (B) and Lmx1a (C). Cultures were processed for the expression of midbrain dopaminergic neuron markers including Nurr1 (D-F), En1 (G-I), and Pitx3 (J-L) in conjunction with TH (inset E, H, K). Nurr1, En1 and Pitx3 was detected in cells derived from PA6 formed progenitors (D, E, G, H, J, K) whereas only Nurr1 (F, M-O) was found to be expressed in cells derived from monolayer-derived neural progenitors. Scalebar for A-C and M-O = 10 μm, scalebar for D, E, G, H, J, K = 50 μm and scalebar for F, I, L = 50 μm.