Fig. 6

Sox32 modulates Oct4 and Nanog complexes in ventrolateral endoderm of gastrula embryos.

(a) The schematic shows the Ventrolateral (VL)- and dorsal (D)-mesendoderm layers at 60% epiboly (7 hpf), with mesoderm in orange and endoderm in yellow. (b, c) Lifetime values (b) and FLIM images (c) of GFP-Oct4 alone and in the presence of mCherry-Nanog in the nuclei of individual cells within VL-Mesoderm and VL-Endoderm cells of wt and ela^br13 mutants. Values of FLIM data represent the median and quartile ranges of data from three to five independent experiments (n = 20–40 cell nuclei from 10 embryos; ***p<0.0001). Scale bar: 5 µm. (d) qRT-PCR analysis of sox32 relative to actin in wt and ela^br13 mutant embryos. Values represent mean ± SEM of data from three independent experiments (**p<0.01). (e) Graphs show percentage of binding of GFP-Oct4 and mCherry-Nanog in VL-Mesoderm and VL-Endoderm of wt and ela^br13 mutant embryos. Values represent the median and quartile ranges from data of three to five independent experiments (n = 20-40 cell nuclei from 7 to 10 embryos; **p<0.01). Values represent mean ± SEM of data from three independent experiments (**p<0.01). n.s. over bars indicates non-significant differences. (f) qRT-PCR analysis relative to actin reveals different transcription levels of bmp2b, bmp4, bmp2a and her5 in ela^br13 mutants at 60% epiboly (7 hpf). Values represent mean ± SEM of data from three independent experiments (**p<0.01). (g) bmp4 expression (top view, dorsal is to the right-hand side) is ventrally reduced in ela^br13 mutants compared with wt embryos. her5 expression (dorsal view) is dorsally upregulated in ela^br13 mutants related to wt embryos. See also Figure 6-figure supplement 1.