Results: 4

1.
Figure 1

Figure 1. Characterization of DILP2 knock-down in UAS-dilp2RNAi/d2GAL flies.. From: Reduction of DILP2 in Drosophila Triages a Metabolic Phenotype from Lifespan Revealing Redundancy and Compensation among DILPs.

The effect of UAS-dilp2RNAi driven by d2GAL in the mNSCs on dilp expression in 7 day old adult female heads was measured by quantitative RT-PCR and Western blot analysis. (A) dilp 2, 3 and 5 relative transcript levels in dilp2RNAi/d2GAL and control female heads. (B) Western blot analysis of DILP2 (top panel) or the tubulin loading control (bottom panel) in total head protein extracted from females of the indicated genotype. (C) dilp 2, 3 and 5 relative transcript levels in d2GAL/UAS-InRDN and control female heads. (D) dilp 2, 3 and 5 relative transcript levels in a FOXO null mutant (FOXO21/25) and control female heads. (A, C and D): data are shown as mean relative expression±SEM (N = 5), * denotes significant difference to controls (P<0.05).

Susan Broughton, et al. PLoS ONE. 2008;3(11):e3721.
2.
Figure 4

Figure 4. The effect of dilp2RNAi expression in mNSCs on hemolymph glucose and trehalose levels, and whole-body trehalose, glycogen and lipid content.. From: Reduction of DILP2 in Drosophila Triages a Metabolic Phenotype from Lifespan Revealing Redundancy and Compensation among DILPs.

(A) Hemolymph glucose and trehalose concentrations in 7 day old mated females, maintained on standard food with 100 g/l of sugar and fasted on 1% agar for 5 hours prior to testing. N = 10 for each genotype. (B) Hemolymph glucose and trehalose concentrations in third instar wandering larvae that developed on standard food with 100 g/l of sugar: UAS-rpr/d2GAL (N = 7), UAS-dilp2RNAiA/d2GAL (N = 7), UAS-dilp2RNAiA/+ (N = 5), UAS-dilp2RNAiB/d2GAL (N = 6), UAS-dilp2RNAiB/+ (N = 6), UAS-rpr/+ (N = 5) and d2GAL/+ (N = 6). (C) Whole-fly trehalose content per mg of fly (fresh weight). UAS-rpr/d2GAL (N = 20), UAS-dilp2RNAiA/d2GAL (N = 20), UAS-dilp2RNAiA/+ (N = 10), UAS-dilp2RNAiB/d2GAL (N = 20), UAS-dilp2RNAiB/+ (N = 10), UAS-rpr/+ (N = 10) and d2GAL/+ (N = 20). (D) Glycogen content per mg of fly (fresh weight), N = 21. (E) Lipid content per mg of fly (fresh weight), N = 17. In all panels, data are shown as mean±SEM, * indicates significant difference to appropriate controls (P<0.05).

Susan Broughton, et al. PLoS ONE. 2008;3(11):e3721.
3.
Figure 2

Figure 2. dilp2RNAi/d2GAL mated females are not long-lived or less fecund than controls.. From: Reduction of DILP2 in Drosophila Triages a Metabolic Phenotype from Lifespan Revealing Redundancy and Compensation among DILPs.

Survival curves, median lifespans, percentage increase compared to control, and sample sizes are as follows. (A): Experiment 1. Median lifespan of: UAS-rpr/d2GAL = 75 days (13.6% increase over UAS-rpr/+ & d2GAL/+ controls, P<0.0001), N = 69; UAS-dilp2RNAiA/d2GAL = 67 days, N = 71; UAS-dilp2RNAiA/+ = 66 days, N = 36; UAS-dilp2RNAiB/d2GAL = 66 days, N = 130; UAS-dilp2RNAiB/+ = 60 days, N = 92; UAS-rpr/+ = 66 days, N = 110; and d2GAL/+ = 66 days, N = 61. (B) Experiment 2. Median lifespan of: UAS-rpr/d2GAL = 81 days (19% increase over UAS-rpr/+ control, P<0.0001), N = 155; UAS-dilp2RNAiA/d2GAL = 76 days, N = 157; UAS-dilp2RNAiA/+ = 73 days, N = 137; UAS-dilp2RNAiB/d2GAL = 71 days, N = 145; UAS-dilp2RNAiB/+ = 69 days, N = 129; UAS-rpr/+ = 68 days, N = 139; and d2GAL/+ = 64 days, N = 153. (C) Experiment 3. Median lifespan of: UAS-rpr/d2GAL = 73 days (12% increase over d2GAL/+ control, P<0.0001), N = 153; UAS-dilp2RNAiA/d2GAL = 68 days, N = 180; UAS-dilp2RNAiA/+ = 67 days, N = 166; UAS-dilp2RNAiB/d2GAL = 65 days, N = 179; UAS-dilp2RNAiB/+ = 67 days, N = 175; UAS-rpr/+ = 59 days, N = 149; and d2GAL/+ = 65 days, N = 169. (D) Fecundity of females from experiment 1 shown in (A). (E) Fecundity of females from experiment 2 shown in (B). Data are shown as mean number of eggs laid per female per day±SEM.

Susan Broughton, et al. PLoS ONE. 2008;3(11):e3721.
4.
Figure 3

Figure 3. Survival of dilp2RNAi/d2GAL female flies under oxidative stress and starvation.. From: Reduction of DILP2 in Drosophila Triages a Metabolic Phenotype from Lifespan Revealing Redundancy and Compensation among DILPs.

Survival curves, median lifespans, percentage increase compared to control, and sample sizes of 7 day old mated females are as follows: (A) Experiment 1 on 5% H2O2. Median lifespans (in days) of: UAS-rpr/d2GAL = 4.04, N = 104, (34% increase over d2GAL/+ control, P<0.0001); UAS-dilp2RNAiA/d2GAL = 3.25, N = 102; UAS-dilp2RNAiA/+ = 3.25, N = 100; UAS-dilp2RNAiB/d2GAL = 3.25, N = 77; UAS-dilp2RNAiB/+ = 2.9, N = 79; UAS-rpr/+ = 3.01, N = 82; and d2GAL/+ = 3.01, N = 98. (B) Experiment 2 on 5% H2O2. Median lifespans (in days) of: UAS-rpr/d2GAL = 3.85, N = 77, (15.6% increase over d2GAL/+ control, P<0.0001); UAS-dilp2RNAiA/d2GAL = 3.21, N = 83; UAS-dilp2RNAiA/+ = 3, N = 96; UAS-dilp2RNAiB/d2GAL = 3.33, N = 58; UAS-dilp2RNAiB/+ = 3.21, N = 84; UAS-rpr/+ = 3.21, N = 91; and d2GAL/+ =  3.33, N = 87. (C–E) Experiment 1 on 1% agar. Median lifespans (in days) of: UAS-rpr/d2GAL (mNSC-ablated) = 5.9, N = 120, (47.5% increase over UAS-rpr/+ & d2GAL/+ controls, P<0.0001); UAS-dilp2RNAiA/d2GAL = 4.17, N = 112, (4.25% increase over UAS-Dilp2RNAiA/+ control, P<0.0001); UAS-dilp2RNAiA/+ = 4, N = 115; UAS-dilp2RNAiB/d2GAL = 4.17, N = 98, (4.25% increase over d2GAL/+ control, P = 0.0004); UAS-dilp2RNAiB/+ = 3.86, N = 117; UAS-rpr/+ = 4, N = 116; and d2GAL/+ = 4, N = 117. (F–H) Experiment 2 on 1% agar, Median lifespans (in days) of: UAS-rpr/d2GAL = 6.2, N = 91, (25% increase over UAS-rpr/+ control, P<0.0001); UAS-Dilp2RNAiA/d2GAL = 4.96, N = 94, (19.5% increase over UAS-dilp2RNAiA/+ control, P = 0.0041); UAS-dilp2RNAiA/+ = 4.15, N = 91; UAS-dilp2RNAiB/d2GAL = 4.15, N = 88; UAS-dilp2RNAiB/+ = 3.96, N = 91; UAS-rpr/+ = 4.96, N = 98; and d2GAL/+ = 3.96, N = 98. (I–K) Experiment 3 on 1% agar, Median lifespans (in days) of: UAS-rpr/d2GAL = 4.85, N = 89, (24% increase over UAS-rpr/+ control, P<0.0001); UAS-dilp2RNAiA/d2GAL = 3.25, N = 85; UAS-dilp2RNAiA/+ = 3.17, N = 94; UAS-dilp2RNAiB/d2GAL = 3.9, N = 76, (23% increase over d2GAL/+ control, P<0.0001); UAS-dilp2RNAiB/+ = 3.06, N = 76; UAS-rpr/+ = 3.9, N = 85; and d2GAL/+ = 3.17, N = 82.

Susan Broughton, et al. PLoS ONE. 2008;3(11):e3721.

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