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Xenbase Image ID: 176004

Figure S1. Metabolic Labeling of Nascent Zygotic Transcripts in Xenopus Embryogenesis using 5-Ethynyl Uridine (EU). Related to Figure 1. (F-G) (F) Image analysis pipeline for quantification of nuclear EU-RNA intensity and amount in wholemount Xenopus embryos at various developmental stages. (G) Heatmap of EU-RNA nascent transcript intensity (top panel) and amount (bottom panel; also shown in Figure 1D) accumulated in the nuclei of individual cells within an embryo during developmental progression. mpf, minutes post fertilization. Embryonic cleavages were approximated from log2 cell number. EU-RNA amount in each nucleus was calculated by multiplying the nucleus volume with its nuclear EU-RNA intensity after background subtraction. The images shown are reconstructed using the x, y and z information of the center of each nucleus from confocal imaging. Color-coded scale indicates the EU-RNA intensity or amount accumulated in the nucleus from low (blue) to high (red). (H) Representative confocal images of EU-RNA (red) and DNA (green) for cells at the animal pole (top panels) and the vegetal pole (bottom panels) in the same embryo at respective cleavage stages. Scale bars, 50 µm. (I) Percentage of transcriptionally active cells within the animal pole (AP, top 200 µm in depth) and vegetal pole (VP, bottom 200 µm in depth) of embryos (N = 3-7 embryos) at indicated times post-fertilization. mpf, minutes post fertilization. Data show mean ± standard deviation. The statistical difference between groups was determined by one-way ANOVA. ****, p < 0.0001.

Image published in: Chen H et al. (2019)

Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V.

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