|
Fig. 6
The state of embryonic RNA granules is changed from solid-like to liquid-like during early stages of development
(A–D) FISH analysis of pou5f3 mRNA (green) in embryos at 0 hpf (A) and 3hpf (C) without (Control) and with hexanediol (Hexanediol) or hexanetriol (Hexanetriol). DNA is shown in blue. Numbers of pou5f3 RNA granules per 3,600 μm2 in embryos at 0 hpf (B) and 3 hpf (D) treated without (−) and with hexanediol (HD) or hexanetriol (HT) were counted (means ± standard deviations; n = 4). Similar results were obtained from two independent experiments. ∗∗p < 0.01, ∗∗∗p < 0.001 (Tukey-Kramer test).
(E) Left, distribution of GFP-Pabc1l in embryos at 3 hpf. Right, time-lapse images of the boxed region in the left image showing fusion of three GFP-Pabp1l spots (arrows) into a larger spherical drop.
(F) Immunoblotting analysis of puromycylated peptides in embryos at 3 hpf treated with (+) or without (−) puromycin (Puro) and hexanediol (HD).
(G) Puro-Pou5f3 PLA sites (red) in embryos at 3 hpf treated without (Control) and with hexanediol (Hexanediol) or hexanetriol (Hexanetriol). DNA is shown in blue. Bars, 20 μm.
(H) Numbers of Puro-Pou5f3 PLA sites per 3,600 μm2 in the embryos treated without (−) and with hexanediol (HD) or hexanetriol (HT) were counted (means ± standard deviations; n = 6). Similar results were obtained from two independent experiments. ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (Tukey-Kramer test).