XB-IMG-173174
Xenbase Image ID: 173174
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Figure 3. (A) Donor nuclei were isolated from oocytes co-expressing GFP-Npm1 and RFP-Fbl, while acceptor nuclei were isolated from un-injected oocytes. (B) Top, soluble protein depleted (SPD) donor nucleus was mixed with oil isolated acceptor nucleus, demonstrating soluble protein depletion does not irreversibly alter nucleoli, and suggests that a soluble nuclear factor(s) is responsible for the dynamic nature of nucleoli. Middle, transfer of labeled protein from oil isolated donor to soluble protein depleted acceptor indicates that soluble protein depletion does not prevent recruitment nor aggregation of soluble proteins. Bottom, aggregated proteins exchange between soluble protein depleted nucleoli. Exposure was adjusted in bottom images to account for decreased signal, and donor nucleolus was included in top inset for comparison. Brightness and contrast were adjusted in dashed inset to aid in RFP-Fbl visualization. Asterisks indicates the acceptor nucleolus. (C) Exchange of fluorescently labeled proteins from oil isolated and merged nuclei singly expressing GFP-Npm1 or RFP-Fbl. (D) Trans-nucleolar fusion following mixing of soluble protein depleted RFP-Fbl expressing donor (red) and oil isolated GFP-Npm1 (green) under oil demonstrates rescue more normal rounded phenotype. (E) Exchange of GFP-Npm1 and RFP-Fbl is diffusion limited. As distance from donor âDâ increases fluorescence intensity of acceptor decreases. Images in (BâC) are representative of 3 independent experiments and at least nine biological replicates (merged nuclei). Images acquisition began 15 min after trans nuclear mixing. The fusion event depicted in (D) is the most complete event we observed. Partial fusions (e.g. middle panel B) are commonly observed. Image published in: Hayes MH et al. (2018) © 2018, Hayes et al. Creative Commons Attribution license Larger Image Printer Friendly View |