Independent replicates) that is a total of 6 fields for two experiments as well as the error bars/standard deviation represent the variation across all six fields and two experiments. Total number of cells scored = 5366, average quantity of cells/field = 894, error bars = s.d. (D) Immunofluorescence analysis of ZsGREEN and TUBB3 expression inside a section of a chick embryo grafted with ZsGREEN+ human axial progenitor-derived trunk NC cells. The DRG region is marked by a yellow box. The photos on the ideal are magnifications of your region marked by the white inset within the DRG area. Arrowheads mark co-localisation in the ZsGREEN and TUBB3 proteins in a donor cell derived, DRG-localised neurite. V, ventral neural tube. Scale bar = one Ethyl acetoacetate Data Sheet hundred mm. (E) Immunofluorescence evaluation of TBX6 (left) or SOX1 (proper) expression in axial progenitors treated with CHIR-FGF2 (pro-PXM circumstances) and RA (pro-PNE conditions) respectively. Scale bar = 100 mm. (F) Top rated left: Representative FACS histogram indicating the gated T-VENUS +hPSC derived axial progenitors at the same time as its flow-sorted fraction (`sort’) which was subsequently plated in NC-inducing conditions. Prime right: Typical percentage of SOX10+ cells (in 5-alpha-reductase Inhibitors Reagents relation to HOXC9 expression) following 5 day differentiation of sorted T-VENUS+ axial progenitors in NC-inducing circumstances, immunostaining and image evaluation. The Figure 3 continued on next pageFrith et al. eLife 2018;7:e35786. DOI: https://doi.org/10.7554/eLife.7 ofResearch post Figure three continuedDevelopmental Biology Stem Cells and Regenerative Medicinedata within the graph were obtained immediately after scoring eight?0 random fields per experiment (N = 5). The error bars/standard deviation represent the variation across all fields and 5 experiments. Error bars = s.d. Bottom: A representative field depicting immunofluorescence analysis of SOX10 and HOXC9 expression in NC cells derived from sorted T-VENUS+ axial progenitors. Scale bar = one hundred mm. (G) qPCR expression evaluation of indicated HOX genes in hPSC-derived anterior cranial (ANC), retinoic acid (RA)-treated NC (+RA), and axial progenitor-derived NC cells (NMP-NC) relative to hPSCs. Error bars = S.E.M. (n = three). (H) qPCR expression evaluation of indicated NC markers in +RA and axial progenitor-derived NC cells relative to untreated anterior cranial NC cells. Error bars = S.E.M. (n = 3). DOI: https://doi.org/10.7554/eLife.35786.008 The following supply data and figure supplements are readily available for figure 3: Supply information 1. Raw data for Figure 3. DOI: https://doi.org/10.7554/eLife.35786.015 Figure supplement 1. Dynamics of trunk neural crest differentiation from axial progenitors. DOI: https://doi.org/10.7554/eLife.35786.009 Figure supplement 1–source data 1. Raw information for Figure 3–figure supplement 1. DOI: https://doi.org/10.7554/eLife.35786.010 Figure supplement two. Characterisation of hPSC- derived axial progenitor differentiation merchandise. DOI: https://doi.org/10.7554/eLife.35786.011 Figure supplement 2–source information 1. Raw information for Figure 3–figure supplement 2. DOI: https://doi.org/10.7554/eLife.35786.012 Figure supplement 3. Quantification of pluripotency marker expression in hPSC-derived axial progenitors. DOI: https://doi.org/10.7554/eLife.35786.013 Figure supplement 3–source data 1. Raw data for Figure 3–figure supplement three. DOI: https://doi.org/10.7554/eLife.35786.Characterisation of axial progenitor-derived trunk NC cellsWe next tested the developmental possible of human axial progenitor-derived trunk NC cells. To.
