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. 2023 Jul 17;33(9):661–678. doi: 10.1038/s41422-023-00846-8

Fig. 6. E-assembloids recapitulate early post-implantation embryogenesis.

Fig. 6

a Schematic representation of improved protocol for assembly and extended culture of E-assembloids. AS, A83-01 + SB431542; CHIR, CHIR99021; LDN, LDN193189-2HCl; ‘+’ and ‘−’ represent ‘add’ and ‘withdraw’, respectively. b Representative phase constrast images of E-assembloids during extended culture. c Representative immunostaining images showing the formation of ACLS and YSLS in D8 E-assembloids. d Quantification of the E-assembloids with EPILCs (red) and XENLCs/ExMLCs (blue) at different time points. n = 3 independent experiments; more than 100 structures were quantified in each experiment; data are presented as mean ± SD. e Representative immunostaining images showing the generation of AVELCs in D8 E-assembloids. f Quantification of the E-assembloids with AVELCs indicated in e. n = 3 independent experiments; more than 100 structures were quantified in each experiment; data are presented as mean ± SD. g Representative immunostaining images showing the specifications of AMELCs (yellow arrowheads) and PGCLCs (white arrowheads) in D8 E-assembloids. h, i Quantification of D8 E-assembloids containing AMELCs and PGCLCs. n = 3 independent experiments; more than 100 structures were quantified in each experiment; data are presented as mean ± SD. AC amniotic cavity, YS yolk sac, LS -like structure. Scale bars, 100 µm. See also Supplementary information, Fig. S6.