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. 2018 Aug 10;7:e35786. doi: 10.7554/eLife.35786

Figure 2. Human axial progenitor cultures exhibit a neural crest/border signature.

(A) Log-fold induction of representative neural crest/neural plate border and BMP-associated transcripts in axial progenitors compared to hESCs. (B) Immunofluorescence analysis of expression of indicated markers in axial progenitors. Magnified regions corresponding to the insets are also shown. Scale bar = 100 µm. (C) Graphs showing the correlation of indicated NC/border markers with T in hPSC-derived (MasterShef7 line) axial progenitors. Each graph corresponds to one scored representative random field and reflects the results from the analysis of 3–5 random fields. Numbers in each quadrant represent percentages of cells belonging to different categories. Numbers of nuclei analysed in each graph: N = 785 (T-SOX9), N = 940 (T-SNAI2), N = 720 (T-PAX3). (D) Immunofluorescence analysis of expression of phosphorylated SMAD1/5 (p-SMAD) in the presence and absence of the BMP inhibitor LDN193189 (LDN). Scale bar = 100 µm. (E) qPCR expression analysis of indicated markers in axial progenitors in the presence (+) or absence (-) of LDN. Error bars = S.D. (n = 3). In all cases nuclei were counterstained with DAPI. PXM, paraxial mesoderm; NC, neural crest. *p<0.05, Paired t-test.

Figure 2—source data 1. Raw data for Figure 2.
elife-35786-fig2-data1.xlsx (11.7KB, xlsx)
DOI: 10.7554/eLife.35786.007

Figure 2.

Figure 2—figure supplement 1. Dissecting the expression of neural crest/border markers in cultures of in vitro-derived axial progenitors.

Figure 2—figure supplement 1.

(A) qPCR expression analysis of indicated NC/border and pluripotency markers in axial progenitors vs hPSCs. Error bars = S.E.M. (n = 3). (B) Quantification of cells marked by different combinations of T-VENUS and indicated NMP/NC-border marker expression in day three axial progenitor cultures derived from a T-VENUS hPSC reporter line (parental line = H9). The data in the graph were obtained after scoring 8–10 random fields per experiment (three independent replicates). The error bars/standard deviation represent the variation across all fields and three experiments. Error bars = s.d. (C) Immunofluorescence analysis of SOX9 expression together with MSGN1-VENUS (top) and TBX6 (bottom) in axial progenitors derived from a MSGN1-VENUS or wild type hPSCs respectively. Scale bar = 100 µm.
Figure 2—figure supplement 1—source data 1. Raw data for Figure 2—figure supplement 1.
elife-35786-fig2-figsupp1-data1.xlsx (11.1KB, xlsx)
DOI: 10.7554/eLife.35786.006