Fig. 2: The deposition of mRNA m5C in oocytes and the in vivo functions of NSUN2 in early embryogenesis. | Nature Communications

Fig. 2: The deposition of mRNA m5C in oocytes and the in vivo functions of NSUN2 in early embryogenesis.

From: Developmental mRNA m5C landscape and regulatory innovations of massive m5C modification of maternal mRNAs in animals

Fig. 2

a Representative images of egg chambers at different stages for D. mel expressing a GFP-NSUN2 fusion protein under the control of its native promoter. DNA was stained with DAPI (blue). Arrows indicate the germ cell that will become the oocyte. The localization of NSUN2 was consistent in all samples of each stage (n = 12, 14, 17, 20). Scale bars, 30 μm. b Representative images of human GV/MI/MII oocytes stained with anti-NSUN2 antibody. 11 GV, 14 MI, and 14 MII oocytes were stained, and the NSUN2 localization was consistent in all oocytes examined. Scale bars, 30 μm. c, d The densities (c) and levels (d) of Type I m5C sites in HeLa cells treated with nocodazole for 0, 24, 48, and 72 h (1 sample per time point). In panel d, a union of sites with levels ≥10% in at least one sample was used for analysis (n = 5039). The density of mRNA m5C sites in MII oocytes is indicated in c. Boxplots: 25th to 75th percentiles (boxes), medians (horizonal lines), and 1.5 times of the interquartile range (whiskers). e Overlaps of Type I mRNA m5C sites between HeLa cells treated with nocodazole for 72 h and MII oocytes. f Comparison of m5C methylation levels in the ovaries between wild-type flies and NSUN2 knockout flies. Type I and Type II m5C sites were shown separately. g Quantification of the percentage of 0–2 h embryos that were in different stages of embryogenesis. Embryos were grouped into four stages: within four cleavage cycles, 5–9 cycles, 10-13 cycles, cellularization (as shown in Supplementary Fig. 9c). The first independent experiment on mutant #1 line was shown. The numbers of flies used are provided in Source Data. h PCA showing the first two PCs, which together explain 82.6% of the variance in the transcriptome data. The amount of variance explained by each PC is indicated on each axis. Wild-type and maternal NSUN2 knockout embryos are colored blue and red, respectively. Source data are provided as a Source Data file.

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