Xenbase Image ID: 176003

Figure S1. Metabolic Labeling of Nascent Zygotic Transcripts in Xenopus Embryogenesis using 5-Ethynyl Uridine (EU). (A-E) Related to Figure 1. (A) EU-microinjection does not interfere with blastula development. (Left) Representative images of buffer and EU-microinjected Xenopus embryos at MBT stage. Embryos at 1-cell stage were microinjected with 10 nl of buffer (1× TBS) and 50 mM EU (final 0.5 mM), respectively. Scale bars, 500 µm. (Right) Quantitation of developmental progression based on surface cell density in buffer and EU-microinjected Xenopus embryos at MBT stage. Cell counts from a 100 × 100-pixel square of the animal pole (N = 10 embryos for each group). Data shown as mean ± standard deviation. Unpaired t test shows no statistically significant difference between buffer and EU-injected embryos. (B) Representative images of nuclei in cells of the animal pole, for embryos at 256-cell stage (log2 cell no. = 8) and cleavage 8000-cell stage (log2 cell no. = 13). EU-RNA (red) and DNA (green). Scale bars, 50 µm. (C) Subcellular localization of nascent RNA. Red, EU-RNA; green, DNA. Scale bar, 10 µm. EU-RNA in the interphase nucleus provides a measure of transcriptional activation within the current cell cycle. Total EU-RNA in a cell indicates the integrated transcriptional history of that cell. Nuclear EU-RNA amount calculated by multiplying the nuclear EU-RNA intensity by nucleus volume. (D) Measurement of EU-RNA intensity in individual embryos as a function of development progression (log2 Cell No.).EU-RNA intensity was measured in EU-microinjected embryos and subtracted with background signal in embryos without EU microinjection. Each red dot represent one embryo. Exponential fitting to the data. Result shows that EU-RNA is exponentially increased strarting from embryonic cleavages 12-13, consistent with previous data. Inset: Measurement of embryo volume as a function of development progression (log2 Cell No.). Each red dot represent one embryo. Linear fitting to the data. Result shows that embryo volume remains relatively constant in early embryogenesis. (E) Measurement of accumulated EU-RNA in embryos from times 380-600 min post-fertilization (log2 cell no. = 11-15). Dot blot assay for biotinylated EU-RNA, using streptavidin-HRP probe. The fold change was normalized to signal in C11; data shown as mean ± standard deviation from triplicates. Image published in: Chen H et al. (2019) Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

Return to previous page