Wild type flies (cantonS) were cultured on standard food (Bloomington stock center, https://bdsc.indiana.edu ) at 20 °C maintaining a 12 h day/night cycle. Eggs from flies kept on respective lipid defined diets were transferred onto fresh lipid defined food and cultivated at 20 °C. Lipid defined food was composed as follows: 1. YAF-Oil: Yeast autolysate (Sigma Aldrich, 100 g/l), Glucose (Roth, 100 g/l), Agar–Agar (Roth, 10 g/l), Nipagin (Sigma Aldrich, 1 g/l) and cold-pressed olive oil (grocery store, 10 g/l). 2. YAF-Sterol: Yeast autolysate (Sigma Aldrich, 110 g/l), Glucose (Roth, 110 g/l), Agar–Agar (Roth, 10 g/l), Nipagin (Sigma Aldrich, 1 g/l) and ß-Sitosterol (Sigma-Aldrich, 1 g/l). Both, YAF-Oil and YAF-Sterol, allow Drosophila to complete multiple regenerative cycles at 20 °C.
Yeast autolysate
Manufactured by Merck Group
Sourced in Germany
Yeast autolysate is a laboratory product derived from the controlled breakdown of yeast cells. It contains a complex mixture of amino acids, peptides, and other cellular components. The core function of yeast autolysate is to provide a nutrient-rich substrate for the growth and cultivation of microorganisms in various experimental and production settings.
Automatically generated - may contain errors
Lab products found in correlation
Glucose, by Carl Roth (2 mentions)
Nipagin, by Merck Group (2 mentions)
Propionic acid, by Nacalai Tesque (1 mentions)
Cholesterol, by Fujifilm (1 mentions)
Butyl p hydroxybenzoate, by Nacalai Tesque (1 mentions)
Glucose, by Merck Group (1 mentions)
Tetracycline, by Merck Group (1 mentions)
Kobe 1 agar, by Carl Roth (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Ampicillin, by Carl Roth (1 mentions)
Ethanol, by Carl Roth (1 mentions)
Agar agar, by Carl Roth (1 mentions)
Sitosterol, by Merck Group (1 mentions)
3 protocols using yeast autolysate
1
Drosophila Lipid-Defined Diet Protocol
2
Cholesterol-Deficient Low-Sterol Food Protocol
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A recipe for Low-sterol food (LSF) was based on Carvalho et al. (2010) (link) with some modifications. LSF used in this study was a mixture of 10% yeast autolysate (Sigma), 10% glucose, 1% purified agar, 0.3% propionic acid, and 0.06% butyl p-hydroxybenzoate (all Nacalai Tesque). cholesterol-supplemented LSF was prepared by adding cholesterol (Wako, Osaka, Japan) to LSF. The final concentration of cholesterol in this food was 6.2 μg/ml. In the original recipe of Eaton’s group, yeast autolysate and agarose for the lipid-depleted medium were chloroform-extracted (Carvalho et al. 2010 (link)). However, we found that even when we did not treat these materials with chloroform, the control larvae on LSF exhibited a reproducible larval-arrest phenotype (Figure 4A ). More importantly, the larval-arrest phenotype on LSF was restored by adding cholesterol to LSF (Figure 4A ), implying that sterol contents in our LSF were significantly reduced. We should note that the phenotype on our LSF was weaker than the phenotype on Eaton’s lipid-depleted medium (Carvalho et al. 2010 (link)), probably because our LSF recipe omitted the chloroform extraction procedure.
3
Longevity Analysis Protocol for Drosophila
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For longevity analysis flies were collected within 24 hours, transferred to fresh vials and left to mate for 48 hours. Female flies were then randomly sorted into vials at a density of 22 individuals per vial. The recipe for lifespan analysis food was modified from Chapman and Partridge [46 (link)]: 7.5% yeast autolysate (Sigma-Aldrich), 7.5% glucose (Roth, Karlsruhe, Germany), 2.1% ethanol (Roth), 2% Kobe I agar (Roth), 0.3% Nipagin (Sigma-Aldrich). To induce geneswitch-gal4 dependent expression, RU486 (Mifepristone, Cayman Chemical Company, Ann Arbor, MI) was added to the food at given concentrations. Antibiotics were applied as a mixture in concentrations of 500 μg/ml ampicillin (Roth), 50 μg/ml tetracycline (Sigma-Aldrich) and 200 μg/ml rifampicin (Sigma-Aldrich) [23 (link)]. Flies were transferred to fresh food every two or three days. Deaths were scored every day.
Calculation of survival, mean lifespan, median lifespan, and Log-rank test was done by Kaplan-Meier analysis using MS Excel and XLSTAT. All values for lifespan experiments are given inS2 Table and S1 –S4 Files.
Calculation of survival, mean lifespan, median lifespan, and Log-rank test was done by Kaplan-Meier analysis using MS Excel and XLSTAT. All values for lifespan experiments are given in
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