Sodium arsenite

For 3-day growth competition experiments with drug-treated cells, we used the following drug concentrations: 2.5 μM sodium arsenite (Ricca Chemical, 714216); 2.5 μM oligomycin A (Santa Cruz Biotechnology, sc-201551); 50 nM rotenone (Sigma-Aldrich, R8875-1G); 10 μM CCCP (Cayman Chemicals, 25458); 5 μM BTdCPU (EMD Millipore, 324892); 10 nM thapsigargin (Sigma-Aldrich, T9033-.5MG); 100 nM tunicamycin (Calbiochem, 65438010); 1.25 μM EN6 (Sigma-Aldrich, SML2689-5MG)^{48 (link)}; 4 nM bafilomycin A1 (Selleck Chemicals, S1413); and 40 nM 17-DMAG (Selleck Chemicals, S1142). For overnight drug treatments, we used 5 μM sodium arsenite, 10 μM CCCP, 0.2 μM oligomycin, 5 μM antimycin A (Santa Cruz Biotechnology, sc-202467) or otherwise indicated in the figure legends. To inhibit the proteasome or autophagy, we used 2 μM carfilzomib (Selleck Chemicals, S2853) for 6 h or 700 nM bafilomycin A1 for 6 h, respectively. ISRIB (Sigma-Aldrich, SML0843) was used at a concentration of 200 nM.

Age-synchronized SD1550 animals were grown to young adulthood on 33-mm NGM plates seeded with OP50 bacteria. Approximately 30 day 0 adults where then picked into 40 μL of either 100 μM hydrogen peroxide (Sigma, UK; 7722-84-1) or 5 mM sodium arsenite (Santa Cruz, DE; 7784-46-5) diluted in M9 for 1- and 2-h exposures, respectively. For rotenone and antimycin A exposures, animals were picked into 40 μL 50 μM or 100 μM concentrations, respectively, diluted in 20 mg/mL OP50/NGM bacteria and incubated for 4 h. Tubes where then washed three times with M9 before animals were pipetted onto slides and immobilized by a coverslip. Both ELT-6 and mitochondrial images were then captured and analyzed as detailed above.