Fig. 4

Telencephalic-like regional identity and neuronal subtype specification of AiNPCs. a Cortex, midbrain, hindbrain, cultured NPCs, and AiNPCs regional-specific gene expression pattern was determined by qPCR. b-e AiNPCs were placed in specific neuronal subtype differentiation media, VGLUT1⁺ glutamatergic neurons (b), GABA⁺ inhibitory neurons (c), Darpp32⁺ inhibitory neurons (d), ChAT⁺ cholinergic neurons (e), and TH⁺ dopaminergic neurons (f) were identified through immunocytochemistry. g Expression of maker genes in AiNPC-derived glutamatergic (VGLUT1), GABAergic (Gad65, Darpp32), cholinergic (Ache) and dopaminergic (Th) neurons was determined by qPCR. h Proportions of neuronal subtypes generated from AiNPCs were determined by immunocytochemistry and shown as a percentage to total neuronal numbers. i-m AiNPCs were placed under mesencephalic cue and glutamatergic/dopaminergic neuron-specific markers and regional-specific gene expressions were determined by immunocytochemistry (i-l) and qPCR analysis (m), respectively. n-p Under the same mesencephalic cue, a dopaminergic neuron-specific marker and regional-specific gene expressions were determined by immunocytochemistry (n, o) and qPCR analysis (p), respectively. Scale bars represent 10 μm (b-f, i-l, n, o). Error bars denote s.d. from triplicate measurements (a, g, h, m, p)