Fig. 5: Induction of posterior neural identity by posterior organizer cells.
From: Changes in cellular composition shape the inductive properties of Hensen’s Node
a Schematic of the grafting experiment: the Hensen′s node from a GFP + donor embryo at stage HH4 or HH6 was transplanted into the marginal zone of a wild-type stage HH4 host. b, c Whole-mount immunostaining for OTX2 (magenta) shows that stage HH4 node grafts induce a secondary axis (arrowhead) that contains anterior neural tissue (pink arrow, n = 5/5 biological replicates with similar results). d, e In contrast, HH6 node grafts induce posterior axial structures (arrowhead), including CHRD-expressing notochord (magenta arrow, n = 5/5 biological replicates with similar results). f Schematic of enhancer reporter experiment: stage HH4 or HH6 donor nodes were grafted into transgenic stage HH4 host embryos, electroporated with SOX2N1::GFP (N1) and SOX2N2::mCherry (N2) reporter constructs. g, h Representative images showing differential enhancer activation following grafting of stage HH4 (g, n = 12/16 biological replicates with similar results) or HH6 (h, n = 11/19 biological replicates with similar results) node tissue. i Box plots showing the distribution of N1-GFP+ and N2-mCherry+ cells per embryo after transplantation of nodes at stages HH4 or HH6. Each dot represents a single embryo (biological replicate); n = 12 embryos (HH4), n = 11 embryos (HH6). Boxes indicate the median (center line), 25th and 75th percentiles (box), and whiskers denote values within 1.5 × interquartile range. A two-way binomial generalized linear model (GLM) was used to assess the effect of stage on marker expression. For GFP+ cells, the stage effect estimate (HH6 vs HH4) was − 1.203 (z = − 9.78, p = 1.5 × 10-22, 95% CI: − 1.444 to − 0.962). For mCherry+ cells, the estimate was + 1.203 (z = 9.78, p = 1.5 × 10-22, 95% CI: 0.962 to 1.444). Source data is provided as a Source Data file.
