TMBIM-2 acts with the calcium pump MCA-3 in regulating cell non-autonomous UPR ^mt activation. (A) Workflow of the IP-MS method in expression TMBIM-2::GFP C. elegans. (B) List of TMBIM-2–interacting proteins that top 10 candidates identified by IP-MS experiments. (C) Interactions of TMBIM-2 with MCA-3. HEK293T cells transfected with the indicated cDNAs and followed by the indicated immunoprecipitations. (D) Representative photomicrographs of ciliated neuronal knockout of mca-3 in neuronal Q40::YFP, dve-1p::dve-1::gfp worms. Scale bar, 250 μm. (E) Quantification of the number of intestinal nuclei puncta with GFP signal per worm as shown in D. n ≥ 15 worms. (F) Fluorescence images of neurons in strains expressing TMBIM-2::GFP with the presence or absence of ciliated neuronal knockout of mca-3. Scale bar = 2 μm. (G) Representative photomicrographs of dve-1p::dve-1::gfp reporter in neuronal Q40::YFP animals with PMCA inhibitor Caloxin 2A1, respectively. Scale bar, 250 μm. (H) Quantification of the number of intestinal nuclei puncta with GFP signal per worm as shown in G. n ≥ 10 worms. (I) Quantification of PM-GCaMP6 maximal fluorescence intensity changes in animals with a vehicle and 10 mM Caloxin 2A1. n ≥ 20 worms. (J) Quantification of the frequency with GCaMP6f fluorescence intensity changes in animals with a vehicle and 10 mM Caloxin 2A1. n ≥ 20 worms. ***P < 0.001; **P < 0.01 via unpaired two-tailed Student’s t test. Error bars, SEM. Source data are available for this figure: SourceData F5.