FGF-signalling in the differentiation of mouse ES cells towards ...

Opposite, Figure 6: In the absence of FGF4, DE rather than VE is formed. Antibody stain and qPCR-analyses ofDE and VE markers in E14, FGF4 +/– and FGF4 –/– cells. A) Undifferentiated cells were stained for SOX17, E-cadherin, SOX7 and the nuclear stain DAPI. B) Cells were differentiated for 5 days in 30 ng/ml activin w/wo 5ng/ml FGF4 and stained for the same markers. Representative images are shown for each condition. Scale bar: 100µm. C) qPCR data showing the relative expression levels of Sox17, Cxcr4, Sox7 and Tdh mRNA present inundifferentiated and differentiated cells of each cell line compared to E14 ESCs, all standardized to the housekeepinggene Tbp. The mean expression ± S.E.M. of 3 independent experiments is shown (n=2 for FGF4 –/– cellstreated w/ activin + FGF4), using a Ratio t-test for the statistical analysis: # = P < 0,05; ## = P < 0,01 compared to theE14 mES cell condition.SUPPLEMENTARY DATAFigure S1: Negative controls of EdU-incorporation and sort gate. A histogram showing the distribution ofpluripotent E14 mES cells with/without EdU-incorporation and without EdU-stain (red and black samples); withoutEdU-incorporation and with EdU-stain (blue sample); and w/ EdU-incorporation and with EdU-stain (stained withAlexa-488, i.e. positive control; green sample).81