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Figure 7. Reprograming cells to a Neural Crest state establishes and requires high MAPK and low PI3K/Akt activity.(A) Western blot of lysates from Pax3-GR/Zic1-GR injected animal pole explants. Explants were cultured alongside sibling embryos and collected at blastula (stage 9) and early neurula (stage 13) stages to examine levels of phosphorylated and unphosphorylated Erk1/2 and Akt. Reprograming to a neural crest state retains the activities of these pathways characteristic of pluripotent blastula cells. (B–C) Animal cap explant assay examining Sox9 and FoxD3 expression in Pax3GR/Zic1-GR injected explants treated with Meki (RDEA119) (B) or co-injected with Act-PI3K (C). Explants were cultured alongside sibling embryos and collected at late neurula stages (stage 18). Blocking MAPK activation or activating PI3K/Akt blocks expression of neural crest markers.Figure 7—figure supplement 1. Reprograming to a Neural Crest state sustains FGFR4 expression.(A) Animal pole explant assay examining FGFR4 expression in animal cap explants injected with Pax3-GR/Zic1-GR. Explants were cultured alongside sibling embryos and collected at blastula (stage 9), midgastrula (stage 12), and midneurula (stage 15) stages. Reprogramming to a neural crest state causes sustained FGFR4 expression. Image published in: Geary L and LaBonne C (2018) © 2018, Geary et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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