Z0152

Cells were exposed to rotenone (Rot, 1 μM; R8875, Sigma), MPP+ (50 μM, DO48, Sigma), zinc (ZnCl₂, 150 μM; Z0152, Sigma), manganese (MnCl₂, 2 μM; 205891000, Acros Organics), and iron (FeCl₃, 1.5 mM; 157740, Sigma) for 24 hours (h). The final concentration of each agent was chosen from a dose response analysis to obtain a submaximal inhibition of cell viability. Prior to stressor addition, cells were pretreated for 1 h with YM-201636 (PIKfyve inhibitor, PIK, 200 nM; 524611, Millipore) or 5-fluoro-2-indolyl des-chlorohalopemide (PLD inhibitor, FIPI, 100 nM; F5807, Sigma) to inhibit the production of PI(3,5)P2 or PA, respectively. To inhibit the proteasome, cells were incubated with MG-132 (100 μM; M7449, Sigma) for 6 h.

We used zinc chloride (ZnCl₂) as a heavy metal stress agent (Sigma-Aldrich catalog # Z0152). We have performed zinc stress experiments in two different life stages: adult flies and embryos.