Figure 4

Derivation of highly enriched SNs without cell sorting. (a) A diagram of the culture conditions for sympathetic neuronal differentiation without cell sorting. (b) FCM analyses of eGFP⁺ cells on days 10, 17 and 31 of the differentiation shown in (a). (c) Quantification of eGFP⁺ cells in (b). Data from three independent experiments are shown in each graph. (d) Immunocytochemistry analyses for eGFP, TH, PRPH and DBH on day 32 of the differentiation (scale bars = 50 µm). (e) Quantification of PRPH⁺, TH⁺ and DBH⁺ cells in (d) (mean ± s.d., n = 3). (f) Immunocytochemistry analyses for PHOX2B and SOX10 on days 10 and 17, and for TH, DBH and PRPH on day 32 (scale bars = 50 µm). The white dashed boxes in the day 10 images mark the areas enlarged in the top left of the same panels. (g) Quantification of PHOX2B⁺ and SOX10⁺ cells on days 10 and 17 in (f) (mean ± s.d., n = 3). (h) Quantification of PHOX2B⁺, TH⁺, DBH⁺ and PRPH⁺ cells on day 32 in (f) (mean ± s.d., n = 3). SB = SB431542, CHIR = CHIR 99021, RA = retinoic acid, DoD = days of differentiation.