Extended Data Fig. 9: Repression of SOX2, Sox3 and CDX2 expression in early-streak progenitors.
(a) Flow cytometry analysis shows that CDX2 expression is uniformly absent from SOX2-OFF and SOX2-ON cultured in FLC medium. (b) Measurement of relative mRNA expression by RT-qPCR shows that early primitive streak markers are induced to comparable levels by the addition of activin (10ng/ml) to FLC medium and in SOX2OFF relative to CEpiLCs. (c) Flow cytometry analysis shows SOX2 levels in EpiLCs, CEpiLCs, pluripotent and early streak progenitors. CHIR concentration was 5μM in 2i+ conditions. (d) Measurement of relative mRNA expression by RT-qPCR shows that Sox3 levels are reduced in early-streak progenitors induced by the addition of activin to FLC medium. (e) Flow cytometry analysis shows the distribution of CDX2 expression is similar, whereas (f) Venus fluorescence is reduced in two Sox2del subclones analysed in Fig. 7f compared to the parental Cdx2 CRE reporter line. (g) Measurement of relative mRNA expression by RT-PCR shows that primitive streak marker expression is reduced in CEpiLCs and SOX2-OFF cultured in FLC medium plus SB-431542. In B, D, and G each data point represents an individual biological replicate. Bars denote mean ± s.e.m. P values calculated for differences of mean expression by two-tailed Student’s t-test are shown. In B for Bra, n = 26 for CEpiLCs and n = 8 for early streak progenitors; for Eomes n = 14 for CEpiLCs and n = 8 for early streak progenitors; for MixL1 n = 14 for CEpiLCs and n = 8 for early streak progenitors; for Nanog n = 14 for CEpiLCs and n = 8 for early streak progenitors. In D n = 5 for CEpiLCs and early streak progenitors. In G for Bra, n = 29 for CEpiLCs, n = 6 for SB treated CEpiLCs, n= 20 for SOX2-OFF, n= 6 for SB treated SOX2-OFF; for Eomes n = 20 for CEpiLCs, n = 6 for SB treated CEpiLCs, n= 12 for SOX2-OFF, n= 6 for SB treated SOX2-OFF; for MixL1 n = 20 for CEpiLCs, n = 6 for SB treated CEpiLCs, n= 12 for SOX2-OFF, n= 6 for SB treated SOX2-OFF; for Nanog n = 20 for CEpiLCs, n = 6 for SB treated CEpiLCs, n= 12 for SOX2-OFF, n= 6 for SB treated SOX2-OFF. Source numerical data are available in source data.
