Extended Data Fig. 5: Elevated ER, osmotic and ROS stress responses in cleavage-resistant fasn-1(D/E) mutant animals.
From: Proteolytic activation of fatty acid synthase signals pan-stress resolution
a, representative images of HSP-4p::GFP stress reporter induction under ER stress in fasn-1(D/E) animals. scale bar, 200μm. b, quantification of HSP-4::GFP stress reporter induction in fasn-1(D/E) animals under ER stress at different time points. Treatment at 32 hr was repeated from main figure (Fig. 2f) for comparison. n = 30 animals. c, quantification of HSP-4::GFP stress reporter induction in a second CRISPR line of fasn-1(D/E) animals under ER stress at different time points. n = 30 animals. d, quantification of fasn-1(D/E) animals escaping larval arrest with tunicamycin (TM) treatment to induce ER stress. n = 5 biological replicates. mean ± SD. e, representative images of NLP-29p::GFP stress reporter induction under osmotic stress in fasn-1(D/E) animals. scale bar, 200μm. f, quantification of NLP-29p::GFP stress reporter induction in fasn-1(D/E) animals under osmotic stress at different time points. Treatment at 32 hr was repeated from main figure (Fig. 2f) for comparison. n = 30 animals. g, quantification of fasn-1(D/E) animals motility with NaCl treatment to induce osmotic stress. n = 3 biological replicates. mean ± SD. h, representative images of GCS-1p::GFP stress reporter induction under ROS stress in fasn-1(D/E) animals. scale bar, 200μm. i, quantification of GCS-1p::GFP stress reporter induction in fasn-1(D/E) animals under ROS stress at different time points. Treatment at 16 hr was repeated from main figure (Fig. 2f) for comparison. n = 30 animals. j, survival for fasn-1(D/E) animals with increasing concentration of PQT to induce ROS stress. n = 5 biological replicates. Mean ± SD, total animals examined, 0 mM, n = 593; 1 mM, n = 604; 2 mM, n = 621; 4 mM, n = 536. *, p < 0.05. Exact p value see Source Data. k, measurement of palmitoleic acid synthesis using 50% deuterated water (D2O) labeling for 16 hours in the presence or absence of ER stress. m + 0, unlabeled palmitoleic acid. m + 1, m + 2, m + 3 are deuterium-labeled representing newly synthesized palmitoleic acid. n = 3 biological samples. Mean ± SD. p values were calculated using Mann-Whitney test (b,f,i) and unpaired t-test (d,g,j).
