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. Author manuscript; available in PMC: 2020 Jul 8.
Published in final edited form as: Nature. 2020 Jan 8;580(7801):113–118. doi: 10.1038/s41586-019-1885-9

Figure 4. FGF signaling regulates the dynamic properties of the segmentation clock.

Figure 4.

a, Left: Scheme illustrating the posterior-to-anterior gradients of FGF and Wnt signaling along the PSM. Right: Immunofluorescence for doubly phosphorylated ERK, β-catenin and DAPI nuclear stain in differentiating human iPSCs and mouse ESCs. n=8 independent experiments. Scale bar = 100μm. b, Average intensity profiles for individual HES7-Achilles human PSM cells treated with vehicle control (DMSO), 250nM PD03 or 250nM PD17. Mean ± 95% CI. n=89 cells (Control), 30 cells (PD03), 34 cells (PD17) c, Average intensity profiles for individual HES7-Achilles human PSM cells treated with vehicle control (DMSO) or 2μM XAV. Mean ± 95% CI. n=67 cells (Control), 29 cells (XAV). d, Number of HES7-Achilles oscillations before arrest in individual human PSM cells treated with 2μM XAV, 250nM PD17 or 100nM/250nM/500nM PD03. Mean ±SD. One-way ANOVA:100nM vs. 250nM p=2.3e−5, 100nM vs. 500nM p=2.2e−10, 250nM vs. 500nM p=1.5e−3. n=34 e, Mean HES7-Achilles intensity for individual HES7-Achilles human PSM cells treated with 2μM XAV, 250nM PD17 or 100nM/250nM/500nM PD03. Mean ±SD. One-way ANOVA: 100nM vs. 250nM p=1.2e−13, 100nM vs. 500nM p=3e−13, 250nM vs. 500nM p=6.9e−6. n=46 (XAV), n=28 (PD17), n=47 (100nM PD03), n=64 (250nM PD03), n=26 (500nM PD03). f, Summary statistics comparing the instantaneous absolute phase difference relative to control for individual cells treated with vehicle control (DMSO), 2μM XAV, 250nM PD17 or 100nM/250nM/500nM PD03. Mean ±SD. One-way ANOVA: control vs. XAV p= 0.0578, control vs. PD17 p=9.2e-8, control vs. 100nM PD03 p=1.3e-14, control vs. 250nM PD03 p=1.1e−8, control vs. 500nM PD03 p=1.1e−5, 100nM vs. 250nM PD03 p=0.8338, 100nM vs. 500nM PD03 p=0.0601, 250nM vs. 500nM p=0.061. n fixed at 11,000 observations. See Extended data Fig. 6n for full histograms. g-i, HES7-Achilles and MESP2-mCherry intensity profiles in small regions of interest within human PSM cultures. Mean ±SD. Dotted line denotes threshold for MESP2 activation (25 a.u.). g, Vehicle control (DMSO) h, 250nM PD03 i, 2μM XAV. n=15.