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. 2018 Mar 22;9:1195. doi: 10.1038/s41467-018-03531-2

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — Nucleolar stress induces dgat-2 expression to promote lipid accumulation. a Confocal microscopy of DGAT-2::GFP in WT, rrp-8(kun54), rsks-1(ok1255), and AD-treated worms. Scale bar represents 20 µm in the panel. b Relative fluorescence intensity of DGAT-2::GFP quantified from a. Data are presented as the means ± SD of at least 20 worms for each worm strain. Significant difference between WT and a specific worm strain, Student’s t-test, ***P < 0.001. c Immunoblotting of DGAT-2::GFP. One-day-old WT, rrp-8(kun54), rsks-1(ok1255), and AD-treated worms expressing DGAT-2::GFP were collected for lysing and immunoblotting with anti-GFP antibody. d Confocal microscopy of DGAT-2::GFP fluorescence in WT, rrp-8(kun54), rsks-1(ok1255), and AD-treated worms treated with either empty vector (EV) or pha-4 RNAi from L1 to day 1 of adulthood. Scale bar represents 20 μm. e Relative fluorescence intensity of DGAT-2::GFP quantified from d. Data are presented as the means ± SD of at least 20 worms for each worm strain. Significant difference between WT and a specific worm strain, Student’s t-test, ***P < 0.001. Significant difference between a specific strain treated with empty vector (EV) and pha-4 RNAi, ^###P < 0.001. f Schematic diagram of the generation of kun140 and kun141 mutations of dgat-2 using multi-sgRNA-directed CRSPR/cas-9 technology (upper panel). Deletion of the kun140 and kun141 mutations of dgat-2 was confirmed using PCR (lower panel). g Nile Red staining of fixed 1-day-old adult worms. Representative animals; the anterior is indicated on the left and the posterior is indicated on the right. Scale bar represents 20 μm. h Distribution of the lipid droplet size (% lipid droplets) measured by Nile Red staining of fixed worms from g. Data are presented as the means ± SD of 10 animals for each worm strain. Significant difference between a specific mutant strain without (EV) and with RNAi treatment, Student’s t-test, ***P < 0.001, **P < 0.01, *P < 0.05. i Percentage of triacylglycerol (% TAG) in total lipids (TAG + phospholipids, PL) analyzed by TLC/GC. Data are presented as the means ± SD of four biological repeats. Significant difference between a specific mutant strain with and without dgat-2 mutant background, Student’s t-test, **P < 0.01, *P < 0.05