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Fig. 3. Evidence for a maternal role for Ascl1 in regulating mesendoderm formation. (A) Schematic of Ascl1-MT reporter construct and the experimental procedures for demonstrating the expression of Ascl1-MT reporter in oocytes and early embryos generated through host transfer. −prog., oocytes without progesterone treatment; +prog. or +p, oocytes treated with 2 µM progesterone. (B) Ascl1-MT reporter expressing oocytes or embryos frozen at the indicated time points after reporter mRNA injection into stage VI oocytes followed by western blotting using an anti-MT antibody. (C,C′) Embryos expressing Ascl1-MT reporter (30 each) at stage 8 separated into animal cap (AC), marginal zone (MZ) and vegetal mass (VM) and frozen immediately after separation followed by anti-MT western blotting. (D) Embryos expressing Ascl1-MT reporter injected with or without Ascl1 MOa at the one-cell stage cultured to stage 6-10 and frozen at the indicated time points followed by anti-MT western blotting. (E) qPCR comparison of gene expression in Ascl1 morphants produced from embryos expressing Ascl1-MT reporter injected with Ascl1 MOa before (oocyte-injection) or after (one-cell stage injection) fertilization. P-values obtained by performing a Student's t-test. (F) Embryos at different stages of development from experiments as described in E. Image published in: Gao L et al. (2016) © 2016. 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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