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. 2021 Oct 8;220(11):e202102122. doi: 10.1083/jcb.202102122

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© 2021 Zhang et al.

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Figure 1. — RNAi screen identified ACS-1 regulating LD size. (A) Candidate genes (2,343) for RNAi screen from 3 previous reports, in which 259 genes were specifically from Maeda (blue), 273 from Kamath (orange), and 392 from Simmer (green), and 1,419 genes were duplicates from 2 or 3 reports (pink). (B) Schematic workflow of RNAi screen in wild-type N2 and transgenic worms for the regulators of LDs visualized by DHS-3::GFP (N2;dhs-3p::dhs-3::gfp) and Nile Red staining of fixed worms. (C) LDs visualized by DHS-3::GFP, fixed Nile Red staining, and LipidTOX staining. DHS-3::GFP worms were imaged by high-resolution laser confocal microscopy; scale bar represents 2.5 and 0.65 µm (enlarged). For Nile Red– and LipidTOX-stained animals, anterior is left and posterior is right. Scale bar represents 5 and 1.25 µm (enlarged). The pink (DHS-3::GFP-positive LDs) and yellow (Nile Red– and LipidTOX-stained particles) arrows indicate represented LDs. (D and E) The size (mean diameter; D) and distribution (percentage; E) of LDs quantified from five representative Nile Red–stained worms for each worm strain. The data are presented as diameter. (F) The abundance of LDs quantified from Nile Red–stained worms. The data are presented as the number of LDs within a fixed area. n, the number of worms scored for quantification. (G) Percentage of TAG in TL (TAG+ PL) by TLC-GC analysis. Data presented are the mean ± SEM of four biological repeats. Significant difference between the control (EV) and a specific RNAi treatment: ***, P < 0.001.