Function of the α/β-hydrolase fold family proteins Pummelig (CG1882) and Hormone-sensitive lipase in the Drosophila melanogaster lipid metabolism

Regulation of Drosophila melanogaster body fat storage by store-operated calcium entry

“…This enzyme hydrolyzes TAG to give rise to DAG and FAs (Grönke et al, 2005). In addition, the Drosophila homolog of mammalian hormone sensitive lipase (Hsl), can translocate to LDs in larvae and adult Drosophila, a process that is likely to be necessary for lipolysis under starvation conditions, although loss-of-function Hsl in adult Drosophila does not prevent normal lipid mobilization during starvation (Grönke et al, 2005;Hehlert, 2016;Kühnlein, 2012). In addition, lipophagy can hydrolyze the lipids of LDs using the autophagy/lysosome system in mammals (Settembre and Ballabio, 2014;Singh et al, 2009).…”

“…For the bicinchoninic acid assay (BCA), flies were homogenized in 600 µL homogenization buffer (0,05% Tween in H2O) and incubated at 70°C (water bath) for 5 min. The detailed procedure is also described in (Hehlert, 2016) Analysis with the above mentioned antibodies was performed successively, i.e. after staining and detection of the antibody reactions, the primary and secondary antibodies were stripped of by incubating the blotted membrane in Restore western blot stripping buffer for 10min at room temperature (Thermofisher Scientific, Cat.…”

“…#: GAGO20-1KT). The procedure was essentially done by following a standard protocol of the laboratory (Baumbach et al, 2014a;Hehlert, 2016;Tennessen et al, 2014). 30 µL of diluted supernatant (1:3-1:5) of the initial homogenate of each sample was measured at 540nm for background absorbance and then mixed with 100 µL of GO Assay reagent in 96-well microtest plate (Sarstedt, Cat.…”

“…TLC involves the differential distribution of molecules in the stationary phase during the capillary force-driven mobile phase migration (Stahl, 1983). TLC of neutral lipids was carried out using the procedures described in Hehlert, 2016). Briefly, 5 adult male flies were homogenized in lipid lysis buffer (150 µL Experiments were repeated twice.…”

“…Lipogenesis assays were done according to the protocol established by (Katewa et al 2012) and (Hehlert, 2016). Briefly, two groups of Stim-TdI 4-5 day old adult male flies were fed with drug-containing Gö-food (RU486 200µM, Sigma-Aldrich, Cat.…”