Extended Data Fig. 6: Characterization of LLPS of LARP-1 with translation machinery proteins and RBPs.
From: Mitochondria-associated condensates maintain mitochondrial homeostasis and promote lifespan
a, Graphic summary of the IDRs in LARP-1, predicted with the Database of Disordered Protein Predictions (D2P2) software. b, Graphic depiction of in vitro LLPS assays. c, Western blotting of LARP-1::GFP in sedimentation assays. S, supernatant, P, pellet. 3 independent experiments with similar results were performed. d, Graphic summary of the correlation of LARP-1::GFP LLPS with NaCl concentration. e, Coomassie-blue staining of the indicated translation proteins and RBPs fused with His6 and Strep tags, as well as mCh or BFP for use in in vitro LLPS assays. ≥ 3 independent experiments with similar results were performed. f, Individual mCh-fused translation machinery proteins (3 μM) as indicated were incubated with GFP or LARP-1::GFP for 1 min or 30 min. Images are shown for mCh and GFP fluorescence. g-h, Time-course images (g) and western blotting (h) of LLPS of LARP-1::GFP with His6-fused PAB-1::mCh, IFE-4::BFP, EIF-4A3::BFP, and RPL-38::BFP. For western blotting, sedimentation was performed and proteins in the supernatant (S) and pellet (P) were detected with antibodies against GFP, mCh and His6. 3 independent experiments with similar results were performed. i-j, Individual mCh-fused RBPs (3 μM) as indicated were incubated with GFP or LARP-1::GFP for 1 min or 30 min. Images are shown for mCh and GFP fluorescence. Scale bars, 5 μm (f-g, i-j).
