All E coli strains carrying the pFVP25 1 plasmid were cultured i

All E. coli strains carrying the pFVP25.1 plasmid were cultured in LB ATM/ATR targets containing 100 μg/mL ampicillin and seeded to NGM plates containing

100 μg/mL ampicillin as described above. Determination of C. elegans total life span and adult life span To determine C. elegans total life span (defined as the number of days from hatching until death), N2, CFC1005 and CFC315 gravid adults were hypochlorite lysed and eggs transferred to NGM plates containing the designated E. coli diet. Two days after hatching coq-3 homozygous mutant worms were HSP inhibitor selected and transferred to plates containing the designated diet. N2 worms were similarly treated. A total of five or six plates per condition were used (20 worms per plate). Worms were scored for survival and moved to new plates every day for the first six days, then every four days thereafter while this website scoring for survival every two days. Worms that responded to being gently prodded with a platinum wire by moving or pharyngeal pumping were counted as alive. Worms with internally hatched larvae, an extruded vulva, or that escaped were censored from the total count. One-way ANOVA analyses of life spans were performed with StatView 5.0.1 (SAS, CA) software at a significance level of 0.05. Similar results were attained when data were subjected to Kaplan-Meier Test at a 0.05 significance level. Maximum life span was calculated from the mean of the top 10% longest lived worms, for each condition. To determine C.

elegans adult life span, N2, CFC315 and EU35 heterozygous gravid adults were hypochlorite lysed and eggs placed on NGM plates containing fresh OP50. After reaching the L4 larval stage, N2, coq-3(ok506) –/ – and skn-1(zu169) –/ – L4 larvae were transferred to separate plates containing either OP50 or GD1 E. coli, and the life span determined as described above. Media swap and UV-treatment of GD1:pAHG E. coli GD1:pAHG and GD1:pBSK cells were grown Uroporphyrinogen III synthase as described above. The cells were pelleted, the spent media was removed

and kept on ice, and the GD1:pBSK cells were discarded. An equal volume of GD1:pAHG cells were resuspended in either their own spent media or the spent media of the GD1:pBSK cells. These suspensions were then seeded onto regular NGM plates, allowed to dry at room temperature, and stored at 4°C until use. Half of the plates containing GD1:pAHG cells in GD1:pAHG spent media and half of the plates containing GD1:pAHG cells in GD1:pBSK spent media were UV-irradiated for 10 minutes at 365 nm on high setting with a Fluorchem2 imaging apparatus (Alpha Innotech, CA). N2 hatchlings were fed OP50 until the L4 larval stage, and then transferred to plates containing one of the designated diets: GD1:pAHG E. coli cells suspended in spent media obtained from cultures of either GD1:pAHG or GD1:pBSK; alternatively these two types of diets were first subjected to UV irradiation prior to the transfer of L4 larvae. Adult life span determinations were performed as described above. Preparation of mixed E.

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