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. Author manuscript; available in PMC: 2016 Aug 10.
Published in final edited form as: Nature. 2016 Feb 10;531(7592):105–109. doi: 10.1038/nature16951

Extended Data Figure 4. Functional characterization of hESC-derived enteric neurons in co-culture with SMCs.

Extended Data Figure 4

a) Schematic illustration of smooth muscle cell (SMC) differentiation protocol. b) Immunofluorescence staining of SMC progenitors for SMA and ISL1. c) Association of various ENC-derived neuron subtypes with SMA+ cells. d) Synapsin::EYFP expression in ENC-derived neurons at 40 days of co-culture with SMCs and stage matched neurons in the absence of SMCs. e) Monoculture of SMCs versus co-cultures of SMCs with ENC-derived neurons. Upper panels: phase contrast images showing morphological changes of SMCs in Co-culture. Lower panels: Immunofluorescence staining of mature SMC markers MYH11 and Acetylcholine Receptor (AchR) in monoculture of SMCs versus co-cultures of SMCs with ENC-derived neurons. f) Diagrams representing extent of contraction of SMC cultures. Arrows indicate the time of pharmacological stimulation. Scale bars= 50 μm in b,c,e; 100 μm in d.