Table 1.
Strains used in this study.
Fig 1.
cup-4 is required specifically in the coelomocytes for axenic lifespan extension.
A. Gene map of cup-4 indicating the deleted regions of alleles used in this study. B. Lifespan in solid fully fed (FF, left) and liquid axenic dietary restriction (ADR, right) conditions of wild-type, cup-4(lst1684) deletion, endogenous cup-4 rescue (cup-4p) and coelomocyte-specific cup-4 rescue (unc-122p) worms. In FF, cup-4(lst1684) (p = 2.0E-15) is long-lived and both endogenous (p = 1.9E-04) and coelomocyte-specific (p = 4.5E-08) rescues are slightly long-lived compared to wild type. In ADR, cup-4(lst1684) is strongly short-lived compared to wild type (p = 5.2E-06) while both endogenous (p = 0.47) and coelomocyte-specific (p = 0.60) rescues do not significantly differ from wild type. C. Bright green fluorescence can be seen in the coelomocytes of a cup-4(syb1015) reporter worm in both FF and ADR (white arrows), alongside wild-type levels of green autofluorescence in the gut. Green autofluorescence is seen in the gut of a wild-type worm in both FF and ADR, although more strongly so in ADR. Scale bar is 250 μm. D. Lifespan in liquid FF and liquid ADR conditions of wild-type, cup-4(ok837) deletion and coelomocyte-ablated worms. In FF, cup-4(ok837) is long-lived (p = 1.6E-10) while the coelomocyte-ablated strain is slightly short-lived (p = 7.1E-06) compared to wild type. In ADR, both cup-4(ok837) (p = 8.0E-16) and coelomocyte-ablated worms (p = 4.0E-16) are short-lived compared to wild type. Mean lifespans, sample numbers and statistical significance are shown in S1 Table.
Fig 2.
daf-16 and pmk-1 are dispensable to axenic lifespan extension.
A. Lifespan in liquid fully fed (FF) and liquid axenic dietary restriction (ADR) conditions of wild-type and of pmk-1(km25), daf-16(jr22), and dbl-1(nk3) deletion worms. pmk-1(km25) does not significantly differ from wild type under FF (p = 0.34), but is short-lived under ADR (p = 1.8E-15). daf-16(jr22) is similarly short-lived under FF (p = 3.6E-15) and ADR (p = 1.2E-15). dbl-1(nk3) is slightly long-lived under FF (p = 0.0059) and slightly short-lived (p = 1.9E-05) under ADR, but an interaction affect with FUdR is likely (S2 Fig). B. Lifespan in liquid FF and liquid ADR conditions of wild-type, pmk-1(km25) deletion, endogenous pmk-1 rescue (pmk-1p), coelomocyte-specific pmk-1 rescue (unc-122p) and intestine-specific pmk-1 rescue (ges-1p) worms. In ADR, pmk-1(km25) is short-lived (p = 1.8E-15) compared to wild type, as are the endogenous (p = 1.7E-10), coelomocyte-specific (p = 1.6E-13) and intestine-specific (p = 9.0E-16) rescues. C. Lifespan in liquid FF and liquid ADR conditions of wild-type and pmk-1(lst1688) deletion worms. No significant difference was observed under FF (p = 0.95) or ADR (p = 0.19). Mean lifespans, sample numbers and statistical significance are shown in S1 Table. D. Lifespan in liquid FF and liquid ADR conditions of wild-type, wild-type pmk-1 in KU25-enriched background, and pmk-1(km25) deletion worms. The worms with a functional pmk-1 in a KU25 background have a similar axenic lifespan to KU25 pmk-1(km25) knockout worms (p = 0.23), while both have a significantly shorter axenic lifespan than wild-type worms (p = 1.2E-15/p = 6.0E-16).