For bafilomycin A1 treatment, semi-intact hearts were incubated with 100 nM of bafilomycin A1 (Fisher Scientific, AAJ61835MCR) in artificial hemolymph (buffer receipt in [22 (link)]) for 2 h at room temperature prior to the appropriate immunostaining. DMSO was used as a control.
For chloroquine treatments, 100 µl of 20 mM chloroquine diphosphate salt, CQ (Fisher Scientific, ICN19391925) was added onto the fly food. Flies were fed with chloroquine for at least 24 h prior to the cardiac analysis or the western blots. To test the toxicity of CQ treatment, we performed a survival analysis by feeding ywR flies with 20 mM CQ and observed the mortality within a period of within two weeks (about 100 flies per treatment). Flies were transferred to fresh CQ food every day and fly mortality was recorded daily.
For chloroquine treatments, 100 µl of 20 mM chloroquine diphosphate salt, CQ (Fisher Scientific, ICN19391925) was added onto the fly food. Flies were fed with chloroquine for at least 24 h prior to the cardiac analysis or the western blots. To test the toxicity of CQ treatment, we performed a survival analysis by feeding ywR flies with 20 mM CQ and observed the mortality within a period of within two weeks (about 100 flies per treatment). Flies were transferred to fresh CQ food every day and fly mortality was recorded daily.
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