Extended Data Fig. 8: Characterization of the colocalization of LARP-1 membraneless organelles with nascent mRNAs and neopeptides of IMMT-1, ATP-2, and RBPs on the mitochondrial surface.
From: Mitochondria-associated condensates maintain mitochondrial homeostasis and promote lifespan
a, Schematic depiction of transgenic expression of 12×SunTag- and 12×MS2-fused immt-1 or atp-2 and the recognition of 12×SunTag in the neopeptides by scFv::mKate2 and MS2 in the nascent mRNAs by MCB::BFP. b-c, Images of foci containing ATP-2neo (recognized by scFv::mKate2, red) and MS2-fused atp-2 nascent mRNA (recognized by MCP-BFP, purple) on the surface of mitochondria in the hypodermis of N2 and larp-1(yq421) animals expressing Psemo-1SunTag::atp-2-MS2, Psemo-1scFv::mKate2, and Psemo-1MCP::bfp. Mitochondria are labeled with TOMM-20::GFP. Boxed regions are magnified in the adjacent “Zoom 2 ×” images. Arrowheads indicate representative foci on mitochondria. ≥150 mitochondria in each animal were analyzed, n = 10 independent animals. d, Colocalization of LARP-1::GFP with foci containing ATP-2neo (scFv::mKate2, red) and MS2-fused atp-2 nascent mRNA (MCP-BFP, purple) in the hypodermis of N2 animals. Boxed regions are magnified in the adjacent Zoom (2 ×) images. e, Quantification of the fluorescence of LARP-1::GFP (green), ATP-2neo (scFv::mKate2, blue) and atp-2 mRNA (MCP::BFP, purple) in the foci along the purple line in the merged image in (d). f, Representative images showing colocalization within foci of LARP-1::GFP and IMMT-1 (scFv::BFP, purple) with the indicated RBPs in the hypodermis of N2 animals. Individual RBPs fused with mKate2 were expressed under the control of the semo-1 promoter with Plarp-1larp-1::gfp, Psemo-1SunTag::immt-1-MS2, Psemo-1scFv::bfp. Scale bars, 5 μm (b, d, f). For statistical analyses, P values were determined using the two-tailed unpaired Student’s t test. Individual data points (mean ± s.e.m.) and exact P values are shown. ***, P < 0.001.
