Figure 6.

Loss of sialylation does not affect primary germ layer formation of mESCs in vitro. A) Principal component analysis of transcriptomic data from undifferentiated (LIF) mESC and after two, four and eight days of EB differentiation of Cmas ^+/+ (green, n=2) and Cmas ^−/− (red and purple, n=3) mESC lines. B)–E) mRNA expression quantified by normalised fluorescence intensity of B) pluripotency factors Oct3/4, Sox2 and Nanog; C) mesodermal markers Brachyury, Fgf8 and Wnt3; D) embryonic ectodermal markers Fgf5 and Otx2; and E) endodermal markers Foxa2 and Sox17 obtained from transcriptome array data. F) Indirect immunofluorescence and immuno‐histochemical analysis of sections from paraffin‐embedded Cmas ^+/+ and Cmas ^−/− EBs after eight days of differentiation. The capacity to form primary germ layers was detected by staining for the marker proteins Disabled‐2, Nestin and α‐smooth muscle actin (α‐SMA).