XB-IMG-171445
Xenbase Image ID: 171445
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Figure 3. FGF signaling directs the transit from a pluripotent to a lineage restricted state through regulation of Erk and Akt activation.(A) Schematic representation of the FGF receptor and select signaling cascades activated downstream, highlighting the Ras/MAPK (Erk) and PI3K/Akt cascades. (B) Western blot of lysates from animal pole explants cultured alongside sibling embryos and collected at blastula (stage 9), midgastrula (stage 11), and early neurula (stage 13) stages to examine levels of phosphorylated and unphosphorylated Erk1/2 and Akt. Pluripotent cells show high pErk while lineage restricted cells display high pAkt. (C) Western blot of lysates from animal pole explants injected with dnFGFR4. 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. Both pErk and pAkt are blocked by dnFGFR4. (D) Animal pole explant assay examining Epidermal Keratin (EPK) and Trim29 expression in explants injected with either dnFGFR4 or dominant- negative PI3K (dnPI3K) or treated with Meki (RDEA119) and collected alongside sibling embryos at early neurula stages (stage 13–14). Meki treatment phenocopies dnFGFR4.Figure 3—figure supplement 1. Meki (RDEA119) and PI3Ki (LY294) block activation of the MAPK and Akt cascades respectively.(A–B) Western blot of lysates from animal pole explants treated with DMSO, Meki, or PI3Ki. Stage 9 (A) or Stage 13 (B) explants were cultured in vehicle or inhibitor treated media and collected after 0, 10, or 20 min to examine levels of phosphorylated and unphosphorylated Erk1/2 and Akt. Meki blocks pErk but not pAkt, while PI3Ki blocks pAkt but not pErk. 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 |