Oxidative and ER Stress Assays in C. elegans

Oxidative and endoplasmic reticulum mediated stress experiments were performed using 200 mM paraquat (unless specified) (Thermo Fisher Scientific, USA) and 25 ng/µl tunicamycin (Sigma-Aldrich, Canada) respectively. Animals were incubated for 1 h, 2 h, 3 h and 4 h, following a previous published protocol^{6 (link)}. The final working concentrations were made in M9. At least 30 animals of each strain were tested in replicates. Means and standard deviations were determined from experiments performed in duplicate. Animals were considered dead if they did not respond to a platinum wire touch and showed no thrashing or swimming movement in M9. Moreover, dead animals usually had an uncurled and straight body shape in comparison to the normal sinusoidal shape of worms.
The electrotaxis assay protocol has been described previously^{55 (link)}. Briefly, synchronized worms were introduced into a microfluidic channel and subjected to an electric field of 3 V/cm. Locomotory data was extracted from recorded videos using custom MATLAB-based worm tracking software. Electrotaxis speed data was plotted using box plots.