FGF-signalling in the differentiation of mouse ES cells towards ...
FGF-signalling in the differentiation of mouse ES cells towards ...
FGF-signalling in the differentiation of mouse ES cells towards ...
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Opposite, Figure 6: In <strong>the</strong> absence <strong>of</strong> <strong>FGF</strong>4, DE ra<strong>the</strong>r than VE is formed. Antibody sta<strong>in</strong> and qPCR-analyses <strong>of</strong>DE and VE markers <strong>in</strong> E14, <strong>FGF</strong>4 +/– and <strong>FGF</strong>4 –/– <strong>cells</strong>. A) Undifferentiated <strong>cells</strong> were sta<strong>in</strong>ed for SOX17, E-cadher<strong>in</strong>, SOX7 and <strong>the</strong> nuclear sta<strong>in</strong> DAPI. B) Cells were differentiated for 5 days <strong>in</strong> 30 ng/ml activ<strong>in</strong> w/wo 5ng/ml <strong>FGF</strong>4 and sta<strong>in</strong>ed for <strong>the</strong> same markers. Representative images are shown for each condition. Scale bar: 100µm. C) qPCR data show<strong>in</strong>g <strong>the</strong> relative expression levels <strong>of</strong> Sox17, Cxcr4, Sox7 and Tdh mRNA present <strong>in</strong>undifferentiated and differentiated <strong>cells</strong> <strong>of</strong> each cell l<strong>in</strong>e compared to E14 <strong>ES</strong>Cs, all standardized to <strong>the</strong> housekeep<strong>in</strong>ggene Tbp. The mean expression ± S.E.M. <strong>of</strong> 3 <strong>in</strong>dependent experiments is shown (n=2 for <strong>FGF</strong>4 –/– <strong>cells</strong>treated w/ activ<strong>in</strong> + <strong>FGF</strong>4), us<strong>in</strong>g a Ratio t-test for <strong>the</strong> statistical analysis: # = P < 0,05; ## = P < 0,01 compared to <strong>the</strong>E14 m<strong>ES</strong> cell condition.SUPPLEMENTARY DATAFigure S1: Negative controls <strong>of</strong> EdU-<strong>in</strong>corporation and sort gate. A histogram show<strong>in</strong>g <strong>the</strong> distribution <strong>of</strong>pluripotent E14 m<strong>ES</strong> <strong>cells</strong> with/without EdU-<strong>in</strong>corporation and without EdU-sta<strong>in</strong> (red and black samples); withoutEdU-<strong>in</strong>corporation and with EdU-sta<strong>in</strong> (blue sample); and w/ EdU-<strong>in</strong>corporation and with EdU-sta<strong>in</strong> (sta<strong>in</strong>ed withAlexa-488, i.e. positive control; green sample).81