Extended Data Fig. 2: NFYB-1 partially regulates UPRmt factors and MCSR (Related to Fig. 2).
From: NFYB-1 regulates mitochondrial function and longevity via lysosomal prosaposin
a–c, nfyb-1(cu13) loss leads to reduction in nuclear localization of pdve-1::dve-1::gfp upon mitochondrial stress induced by exposure to cco-1i (pointed by white arrow), AL day 1 adults a, microscopy images scale bar = 10 μm, b, quantitation of pdve-1::dve-1::gfp expression using biosorter (n ≥ 400) and c, representative immunoblot of pdve-1::dve-1::gfp, with quantitation of DVE-1::GFP and histone loading control (N = 4, n = 100 per repeat, per condition). d, Kaplan–Meier survival curve of isp-1(qm150) and isp-1(qm150); nfyb-1(cu13) shows that dve-1i does not further reduce isp-1;nfyb-1 lifespan (n = 120 per repeat, per condition). e, nfyb-1(cu13) mutation reduces nuclear localization of patfs-1::atfs-1::gfp upon mitochondrial stress induced by hsp-6i, L3-L4 larvae, scale bar = 10 μm (n ~ 30). f, nfyb-1(cu13) shows no significant (ns) effect on publ-5::gfp expression upon exposure to cco-1i or control luci, day 1 adult (n ≥ 400). g, nfyb-1(cu13) loss only partially reduces phsp-6::gfp expression upon cco-1i induced mitochondrial stress, day 1 adult (n ≥ 400). h, i nfyb-1(cu13) reduces phsp-16::gfp expression upon exposure to hsp-6i from day 1 to day 3, showing a requirement for MCSR. h, microscopy images, scale bar = 75 μm, i, quantitation of phsp-16::gfp expression using biosorter (n ≥ 200). j, k, nfyb-1(cu13) leads to a reduction in lipids by BODIPY staining from day 1 to day 3. j, microscopy images, scale bar = 10 μm, k, quantitation of expression using biosorter (n ≥ 200). a-k, under all WT conditions, cco-1i and hsp-6i induce the mitochondrial stress response. b, d, e-g, i, k All data represent N = 3 independent biological replicates, n = total worms per condition from three replicates unless stated otherwise. Error bar shows mean ± s.e.m, b, e-g, i, k statistics determined by one-way ANOVA and c, two-sided t-test. *P < 0.5, **P < 0.01, ***P < 0.001. d, Two-sided Mantel–Cox log-rank test, refer to Supplementary Table 2 for statistics.
