C. elegans thrashing assays

Experiments were performed on age synchronized worms by picking L4 stage a day before the assay. The next day, one-day old adult hermaphrodite C. elegans were picked into 900 μL of M9 buffer containing 0.1% BSA in a 12 well plate. The worms were left to settle for 10 min. The number of thrashes was counted for 1 min. Then 100 μL of either M9 (control), Lev or Pyr solution was added to the wells to give a final concentration of 10, 25, 50, 100 or 150μM Lev or Pyr. The number of thrashes was counted after 10, 20, 30 and 40 min (and 60 min for some concentrations) of exposure to Lev. The data were plotted as a mean ± standard error of the mean.