Figure 1. NHR-49 is essential for the longevity of germline-depleted animals and is widely expressed in somatic cells. A: Effect ofnhr-49 RNAi on the lifespan of germline defective
glp-1 adults.
glp-1 mutants were subjected to RNAi during adulthood by feeding bacteriacontaining control (empty) vector (green; m = 26.360.6, n = 92/96) as well as bacteria expressing dsRNA targeting
daf-16 (blue; m = 16.460.2, n = 92/97; P vs. control ,0.0001),
nhr-49 RNAi clone #1 (red; m = 17.360.3, n = 99/101; P vs. control ,0.0001, P vs.
daf-16 RNAi 0.01) and
nhr-49 RNAi clone#2 (maroon; m = 15.560.1, n = 79/92, P vs. control, ,0.0001, P vs.
daf-16 RNAi 0.005). Clones #1 and #2 were obtained from the Ahringer and Vidalfeeding RNAi libraries [39,40], respectively. B: Effect of
nhr-49 mutation on the lifespan of
glp-1 mutants and wild-type (N2) worms.
glp-1(green; m = 31.060.5, n = 94/101),
nhr-49;
glp-1 (red; m = 14.160.1, n = 95/97; P vs.
glp-1,0.0001), N2 (black; m = 21.660.1, n = 75/98),
nhr-49 (brown;m = 14.460.2, n = 89/100; P vs. N2,0.0001). C: Effect of
nhr-49 mutation on the lifespan of
daf-2 mutants. N2 (black; m = 16.960.1, n = 48/80),
nhr-49 (brown; m = 10.760.1, n = 80/91; P vs. N2,0.0001).
daf-2(
e1368) (blue; m = 30.660.4, n = 31/75; P vs. N2,0.0001),
nhr-49(
nr2041);
daf-2(
e1368)(pink; m = 29.560.7, n = 45/86; P vs. N2,0.0001; P vs.
daf-2(
e1368) 0.73).
daf-2(
e1370) (green; m = 43.060.6, n = 42/67; P vs. N2,0.0001),
nhr49(nr2041);
daf-2(
e1370)(red; m = 42.460.6, n = 46/74; P vs. N2,0.0001; P vs.
daf-2(
e1370) 0.004). D: Effect of
cyc-1 RNAi on lifespan of N2 andnhr-49. N2 worms grown on control vector bacteria (black; m = 16.960.3, n = 81/90) and on
cyc-1 RNAi bacteria (green; m = 19.660.3; n = 85/90; P vs.control ,0.0002; percent increase in lifespan: 14).
nhr-49 mutants grown on control vector bacteria (purple; m = 11.660.1; n = 86/89) and on cyc-1RNAi bacteria (red; m = 14.860.2; n = 85/95; P vs. control ,0.0001; percent increase in lifespan: 22). E-H: NHR-49::GFP expression in adultsomatic tissues. NHR-49::GFP is visible in the cytoplasm and nuclei of neurons (E), muscle (F), hypodermis (G) and intestinal cells (H). Pmyo2::mCherry,the co-injection marker, is seen as red fluorescence in the pharynx in E. I: Rescue of the shortened lifespans of
nhr-49 and
nhr49;
glp-1mutants by the NHR-49::GFP fusion protein. N2 (black; m = 22.860.2, n = 81/92),
glp-1 (green; m = 36.360.2, n = 74/104, P vs. N2,0.0001),
nhr-49;
glp-1 (brown; m = 17.960.2, n = 104/106, P vs.
glp-1,0.0001),
nhr-49;
glp-1;NHR-49::GFP non-transgenic siblings (purple; m = 18.460.2,n = 101/107, P vs.
glp-1,0.0001, P vs.
nhr-49;
glp-1 0.28),
nhr-49;
glp-1;NHR-49::GFP (red; m = 35.660.4, n = 58/102, P vs.
glp-1 0.95, P vs.
nhr-49;
glp-1,0.0001, P vs. non-transgenic siblings ,0.0001). All lifespan data are shown as mean lifespan in days (m) 6 standard error of the mean (SEM). 'n' refers to the number of worms analyzed divided by total number of worms tested in the experiment (some worms were censored from the analysis asdescribed in the methods section). P values were calculated using the log rank (Mantel Cox) method. Data from additional trials of these experimentsare presented in S2 (panel A), S3 (panels B and I), S4 (panel C and D) Tables.