Figure 2

SOX10 and PHOX2B, essential transcription factors of sympathetic NCCs, are expressed in CD49d⁺eGFP⁺ cells. (a) A diagram of the culture condition for sympathetic NCC differentiation. (b) Representative images of day 10 aggregates of KhES1 control and KhES1 PHOX2B::eGFP (scale bars = 200 µm). (c) Immunocytochemistry analyses of unsorted and CD49d⁺eGFP⁺-sorted day 10 differentiated cells for PHOX2B and SOX10 (scale bars = 50 µm). The white dashed boxes mark the areas enlarged in the right panels. The white arrowheads indicate PHOX2B⁺SOX10⁻ cells. (d) Quantification of PHOX2B⁺ and SOX10⁺ cells in (c) (mean ± s.d., n = 3). (e) Immunocytochemistry analyses for HOXB7 on day 0 and day 10 (CD49d⁺eGFP⁺-sorted) of differentiation (scale bars = 50 µm). (f) Quantification of HOXB7⁺ cells in (e) (mean ± s.d., n = 3). SB = SB431542, CHIR = CHIR 99021, RA = retinoic acid.