ATP determination in live worms

For determination of ATP concentrations, approximately 2,000 PE327 L1 larvae [64] were placed onto NGM plates seeded with either E. coli OP50 or E. coli HT115 and allowed to develop to young adult stage at 25°C. Worms were then transferred to 96-well, white clear bottom plates and washed with S-basal five times. 150 μL Luminescence buffer (0.14M K2PO4, 0.03M sodium citrate, 1% DMSO, 1mM luciferin) was added to each well in the plate. Bioluminescence and GFP fluorescence were measured after 30 min using a Cytation5 multimode plate reader/imager (BioTek). Luminescence values were normalized to GFP fluorescence to control for differences in protein expression.