Supplement 1. Primers for RT-PCR. PCR was performed using the primers indicated as below under optimal amplification conditions (95 °C for 5 min; 23-35 cycles of 95 °C for 1 min, 52 °C for 30 sec, 72 °C for 1.5 min; 72 °C for 7 min) for each gene. The PCR amplification of each cDNA was performed independently using three samples in triplicate.

Gene / Forward / Reverse
daf-16 / CATGGATCCATCCAGATGCAAAGCCAG / CATGGATCCGTATGCTGTGCAGCTACA
sir-2.1 / CATGGATCCATGTCACGTGATAGTGGC / CATGGATCCCGACCAGAAGTTTCAAGC
nmnat-1 (F26H9.4) / CATGGTACCTGAAACGAGTCGCTCTTC / CATGGTACCTAACCTCATCTGGCGTCG
nmnat-2 (W06B3.1) / CATGAATTCAGTCGGGTCATTCAATCC / CATGAATTCTTACGGAATCTGGTGTGG

Supplement 2. Primers for RT-PCR. PCR was performed using the primers indicated as below under optimal amplification conditions (95 °C for 5 min; 25-30 cycles of 95 °C for 1 min, 52 °C for 30 sec, 72 °C for 1.5 min; 72 °C for 7 min) for each gene. The PCR amplification of each cDNA was performed independently using three samples in triplicate.

Gene / Forward / Reverse
sod-3 / TGCTGCAATCTACTGCTC / ATCCTGGTTTGCACAGGT
sod-4 / TAATTCTGGCTCTCTCCG / GACGGTACCGATAGTTCC
fat-7 / CATGGATCCCTCAATACCTGTTCATGG / CATGGATCCCAGCATCTGTATCAGTCC
gpd-1 / ATGTCGAAGGCCAACGTC / TCGCCAGTGGTGCCAGAC

Supplement 3. Primers for RT-PCR. PCR was performed using the primers indicated as below under optimal amplification conditions (95 °C for 5 min; 25 cycles of 95 °C for 1 min, 52 °C for 30 sec, 72 °C for 1.5 min; 72 °C for 7 min).

Gene / Forward / Reverse
sod-g3 / CATGGATCCTGCAGTGATTCAGAGAGG / CATGGTACCGCACAGGTGGCGATCTTC

Supplement 4. The detailed data of each graph.

Fig. 1A

NAD / Number of worms (n) / Trial / Mean lifespan ± SEM (day) / Significance to control (%)
(-) / 151 / 3 / 20.10±0.59 / 100
10 M / 149 / 3 / 22.71±0.59 / 113
100 M / 145 / 3 / 23.15±0.61 / 115
100 M / 128 / 3 / 22.28±0.63 / 111

Fig. 1B

Number of worms (n) / Trial / Mean lifespan ± SEM (day) / Significance to control (%)
control (L4440) / 135 / 3 / 22.50±0.53 / 100
control+NAD100 μM / 139 / 3 / 25.40±0.63 / 113
sir-2.1RNAi / 138 / 3 / 22.73±0.43 / 97
sir-2.1RNAi+NAD100 μM / 135 / 3 / 22.87±0.60 / 98
sir-2.1 (ok434) / 136 / 3 / 22.21±0.53 / 99
sir-2.1 (ok434)+ NAD100 μM / 137 / 3 / 22.45±0.55 / 100

Supplement 5. The detailed data of each graph.

Fig. 2A

Number of worms (n) / Trial / Mean body size (mm) / Standard error
control (L4440) / 239 / 3 / 1.19 / 0.0080
control+NAD100 μM / 260 / 3 / 1.23 / 0.0089
control+NAD100 μM / 307 / 3 / 1.28 / 0.0095
control+NAD100 μM / 230 / 3 / 1.23 / 0.0085
eat-2 (ad1116) / 209 / 3 / 0.65 / 0.021

Fig. 2B

WT / 0 day / 1 / 2 / 2 / WT+NAD10μM / 0 day / 1 / 2 / 3
Mean number of eggs / 47.1 / 108.8 / 15.2 / 0 / Mean number of eggs / 59.2 / 118 / 14.6 / 0
Standard error / 3.93 / 17.36 / 3.15 / 0 / Standard error / 1.66 / 13.1 / 4.45 / 0
Standard deviation / 14.7 / 64.96 / 11.7 / 0 / Standard deviation / 3.7 / 29.5 / 9.94 / 0
Number of worms (n) / 14 / 14 / 14 / 14 / Number of worms (n) / 15 / 15 / 15 / 15
Trial / 3 / 3 / 3 / 3 / Trial / 3 / 3 / 3 / 3
WT+NAD100 μM / 0 day / 1 / 2 / 3 / WT+NAD1000 μM / 0 day / 1 / 2 / 3
Mean number of eggs / 43.8 / 106.2 / 33 / 4.53 / Mean number of eggs / 43.2 / 101 / 32.7 / 6.43
Standard error / 4.38 / 8.52 / 5.74 / 2.62 / Standard error / 4.72 / 10.4 / 7.07 / 4.42
Standard deviation / 16.9 / 33 / 22.2 / 10.1 / Standard deviation / 17.6 / 38.9 / 26.4 / 16.5
Number of worms (n) / 15 / 15 / 15 / 15 / Number of worms (n) / 14 / 14 / 14 / 14
Trial / 3 / 3 / 3 / 3 / Trial / 3 / 3 / 3 / 3
eat-2 (ad116) / 0 day / 1 / 2 / 3
Mean number of eggs / 8.43 / 15 / 14.1 / 2.75
Standard error / 2.13 / 4.53 / 3.74 / 0.90
Standard deviation / 8.51 / 18.12 / 14.9 / 3.62
Number of worms (n) / 15 / 15 / 15 / 15
Trial / 3 / 3 / 3 / 3

Supplement 6. The detailed data of each graph.

Fig. 3A

NAD / Number of worms (n) / Trial / Mean lifespan ± SEM (day) / Significance to eat-2 (%)
WT / 60 / 3 / 22.50±0.53 / 71
eat-2(ad1116) / 63 / 3 / 31.50±0.94 / 100
eat-2(ad1116)+NAD100μM / 60 / 3 / 34.54±1.02 / 110
eat-2(ad1116)+NAD1000μM / 58 / 3 / 35.60±1.24 / 113

Fig. 3B

Number of worms (n) / Trial / Mean lifespan ± SEM (day) / Significance to control (%)
Control (L4440) / 130 / 3 / 22.40±0.43 / 100
control+NAD100μM / 135 / 3 / 25.82±0.53 / 115
daf-16RNAi / 138 / 3 / 17.20±0.49 / 77
daf-16RNAi+NAD100μM / 131 / 3 / 17.60±0.40 / 79
daf-16(mgDf50) / 135 / 3 / 17.58±0.53 / 78
daf-16(mgDf50)+NAD100μM / 135 / 3 / 17.38±0.52 / 78

Supplement 7. The detailed data of the graph.

Fig. 4B

Number of worms (n) / Trial / Mean survival ± SEM (%)
WT / 60 / 3 / 58.33 ± 3.3
WT+NAD 100μM / 58 / 3 / 85.11 ± 6.1
daf-16(mgDf50) / 55 / 3 / 17.39 ± 3.6
daf-16(mgDf50)+NAD 100μM / 60 / 3 / 13.33 ± 3.3
sir-2.1(ok434) / 60 / 3 / 39.29 ± 5.3
sir-2.1(ok434)+NAD 100μM / 58 / 3 / 44.83 ± 5.4
NADH 100μM / 60 / 3 / 60.42 ± 7.3

Supplement 8. The detailed data of each graph.

Fig. 6A

Number of worms (n) / Trial / Mean lifespan ± SEM (day) / Significance to control (%)
Control (L4440) / 45 / 3 / 24.34±0.55 / 100
nmnat RNAi / 45 / 3 / 21.28±0.55 / 87

Fig. 6B

Number of worms (n) / Trial / Mean lifespan ± SEM (day)
WT / 60 / 3 / 58.33 ±5.89
nmnat RNAi / 58 / 3 / 60.41 ±7.66

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