Science - USA (2022-06-10)

Acosta-Rodríguezet al., Science 376 , 1192–1202 (2022) 10 June 2022 6of11

A

C

Aging_DE

Up

Down

Same

AL

14%

4%

CR.spread

7%

3%

CR.day.2h

7%

3%

CR.day.12h

6%

2%

CR.night.2h

2%

2%

CR.night.12h

5%

3%

B

71%

29%

Up/D

ow

n

NS

Any Feeding

AL down (339)

6 (25%)

21 (7%)

25 (7%)

19 (9%)

6 (43%)

12 (17%)

46 (6%)

46 (6%)

46 (6%)

21 (13%)

aromatic amino acid...

small molecule catabolic...

reg of hormone levels

organic acid catabolic...

homocysteine metab...

α−amino acid catabolic ...

organic acid metab...

oxoacid metab...

carboxylic acid metab...

α−amino acid metab ...

04

−log10(adjp)

63 total terms

GO:BP

AL up (894)

53 (10%)

104 (10%)

113 (10%)

70 (12%)

69 (13%)

179 (10%)

86 (12%)

70 (13%)

68 (14%)

100 (13%)

innate immune resp...

resp to external biotic...

defense response

lymphocyte activation

inflammatory response

immune system process

leukocyte activation

reg of cytokine prod...

reg of cell activation

cell activation

08

−log10(adjp)

948 total terms

D

E

8 (2%)

9 (2%)

11 (2%)

3 (15%)

14 (3%)

23 (2%)

24 (2%)

adaptive immune res..

T cell activation

lymphocyte activ...

antigen processing

innate immune res..

immune response

resp to external bio...

02 4 6

−log10(adjp)

GO:BP

CR

protection

Lifespan

+ Fasting

(day or night)

Lifespan

+ Circadian

alignment

Lifespan

0

1

2

0

1

2

0

1

2

−3

−2

−1

0

1

0

1

2

3

−1

0

AL

CR.night 12h

CR.night 2h

CR.day 12h

CR.day 2h

Aging LFC CR.spread

0

1

2

0

1

0

1

2

0

1

Aging LFC

AL CR.spread CR.day 2h CR.night 12h CR.night 2hCR.day 12h

0

1

−1

0

−2

−1

0

Aging LFC^0

1

2

AL CR.spread CR.day 2h CR.night 12h CR.night 2hCR.day 12h

−6

0

6

−6 −6 0 6

0

6

−6 0 6 −6

0

6

−6 −6 0 6

0

6

−6 −6 0 6

0

6

−6 0 6

Log2FC CR

Log2 FC_AL

R = 0.44

p = 2.2 e-16

R = 0.55

p = 2.2 e-16

R = 0.46

p = 2.2 e-16

R = 0.47

p = 2.2 e-16

R = 0.11

p = 2.2 e-16

CR.spread CR.day.2hCR.day.12h CR.night.2hCR.night.12h

100 total terms

10 (9%)

cold−induced thermog... 8 (7%)

reg of cold−induced... 8 (7%)

temp homeostasis

adaptive thermogenesis

0 12

−log10(adjp)

10 (7%)

4 total terms

GO:BP

Fig. 4. CR ameliorates age-related changes in liver gene expression
observed under AL conditions.(A) Schematic comparison of differential
gene expression between young and old mice in the six feeding conditions.
Circles show the percentage of genes that are unchanged between young
and aged mice (gray), increased in aged mice (red), and decreased in aged
mice (blue). Pie chart on the right shows the percentage of genes susceptible
for age-related changes in any feeding condition. (B) Spearman correlation
plots comparing changes in gene expression between the aging DE genes
between AL and CR groups (aging DE genes are defined here as the
4077 genes that change with age in any of the six feeding conditions tested).

(C) Schematic representation of genes that are protected from age-related

changes in every CR group (left) with GO terms of those significantly up-

regulated (middle) or down-regulated (right) in aged AL mice. Represented

are 10 nonredundant of the top 25 most significant enriched terms. Examples

of age-related fold changes (log 2 FC ± SE) in gene expression are shown

below for all feeding conditions. Gray-shaded areas indicate FC < 1.5

considered as not significant change. (D andE) Schematic representation,

GO, and representative genes that maintain similar levels between young

and old ages due to fasting (day or night) (D) and circadian alignment of

feeding and fasting cycles (E).

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