Journal: bioRxiv
Article Title: Two sequential waves of mRNA translation drive embryonic development
doi: 10.1101/2025.09.18.676998
Figure Lengend Snippet: a Double staining of GFP-Ewsr1b (green) and GFP-Ewsr1b mRNA (red) in embryos injected with GFP-Ewsr1b mRNA carrying Long-3′UTR (Long) or Short-3′UTR (Short) at 4 hpf. Scale bars: 20 µm. b Violin plots showing distances from the nuclear center to signals of GFP-Ewsr1b mRNA carrying Long-3′UTR or Short-3′UTR (means ± SD; n = 80). Similar results were obtained from two independent experiments. **********p < 0.0000000001 (Student’s t -test). c Immunofluorescence of Importin β1 in embryos at 3 hpf. d Immunoblotting of embryos at 3 hpf following IP with control IgG (IgG) or anti-Importin β1 (α-Im β) antibody, and RT-PCR for ewsr1b -3′Long and α-tubulin mRNAs. Similar results were obtained from two independent experiments. e Double staining of Importin β1 (red) and the ewsr1b -3′Long mRNA 3′UTR (green) in embryos at 3 hpf. Left: High-resolution confocal image; right: enlarged views of the boxed area. Similar results were obtained from two independent experiments. f Immunofluorescence of Importin β1 and Ewsr1b in uninjected embryos (Control) or embryos injected with Importazole at 3 hpf. DNA is shown in blue. Enlarged views of the boxed area with or without DNA staining are shown on the right side. Scale bars, 10 µm. g Quantification of average signal intensity in the nucleus per 25 µm 2 . (means ± SD; n = 10). ***********p < 0.00000000001 (Student’s t -test).
Article Snippet: Proteins were separated by SDS-PAGE, transferred onto Immobilon membranes, and probed with primary antibodies; mouse anti-Syncrip antibody (1:1,000, hnRNP Q; Santa Cruz Biotechnology, I8E4; sc-56703), rabbit anti-Syncrip antibody (1:1,000, Proteintech, 14024-1-AP), rabbit anti-Rpl11 antibody (1:1,000, Abcam, ab79352), rabbit anti-Pou5f3 antibody (1:100), mouse anti-GFP antibody (1:1,000, Roche, 11814460001), mouse anti-Ewsr1b antibody (1:100, present study), rabbit anti-Importin β1 antibody (1:1,000, Proteintech, 10077-1-AP), and mouse anti-HuR antibody (1:1000, Santa Cruz Biotechnology; sc-5261).
Techniques: Double Staining, Injection, Immunofluorescence, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Staining
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