Fig. 4. The life-span extension of a lin-14(lf) mutant is daf-16 dependent. (A) The life-span extension conferred by the lin-14(n179)lf mutation is abolished with daf-16(RNAi) (light green); daf-16(RNAi) animals (green—obscured by light green). lin-14(lf); daf-2(RNAi) (light blue) displays a further lengthening of the life-span extension conferred by daf-2(RNAi) (blue) at 25°C. Wild-type (red) and lin-14(n179)lf (pink) animals on mock RNAi are shown for comparison. N2: n = 60, m = 8.9. lin-14(n179)lf: n = 56, m = 11.9, P < 0.0001*. daf-2(RNAi): n = 58, m = 17.1. lin-14(n179)lf; daf-2(RNAi): n = 56, m = 23.0, P < 0.0001*. daf-16(RNAi): n = 55, m = 8.3. lin-14(n179)lf; daf-16(RNAi): n = 54, m = 8.4, P = 0.9330*. (B) lin-14(RNAi) is unable to extend the life span of daf-16(mu86) (light blue) or daf-2(e1370)lf (light green) mutants as compared with these strains grown on mock RNAi (blue and green, respectively) at 15°C. Wild-type animals on mock RNAi (red) and on lin-14(RNAi) (pink) are shown for comparison. N2: n = 45, m = 21.6. lin-14(RNAi): n = 51, m = 25.1, P = 0.0003*. daf-2(e1370)lf: n = 49, m = 32.3. daf-2(e1370)lf; lin-14(RNAi): n = 53, m = 31.1, P = 0.0100#. daf-16(mu86): n = 28, m = 16.4. daf-16(mu86); lin-14(RNAi): n = 42, m = 16.7, P = 0.2300#. (C) A wild-type copy of lin-4 is required for full life-span extension by daf-2(RNAi) (blue versus light blue), and is also required for the life-span phenotype conferred by daf-16(RNAi) (purple versus light purple). Wild-type (red) and lin-4(e912)lf (pink) animals grown on mock RNAi are shown for comparison. N2: n = 70, m = 16.0. lin-4(e912)lf: n = 52, m = 6.8, P < 0.0001*. daf-2(e1370): n = 67, m = 25.5. lin-4(e912)lf; daf-2(RNAi): n = 55, m = 13.8, P < 0.0001*. daf-16(RNAi): n = 64, m = 10.7. lin-4(e912)lf; daf-16(RNAi): n = 55, m = 6.1, P < 0.0001*. All experiments were repeated at least once with similar results. P* and P# are defined in the legend to Fig. 1.
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