Permeabilized C. elegans Embryo Drug Treatment

Drug treatment of C. elegans embryos was performed on permeabilized eggs. For this, young adult hermaphrodites were injected with perm-1 dsRNA and incubated at 20°C for 16 h after the injection. perm-1(RNAi) embryos were than extruded from the gravid hermaphrodite in a solution of egg buffer or 20 mM arsenite (MerckMillipore) diluted in egg buffer or 20 mM arsenite plus 250 μg/ml cycloheximide (Sigma Life Sciences, C7698-1G; Lee et al., 2020 (link)) diluted in egg buffer on a 22×40 mm coverslips. The coverslips with dissected worms and embryos were incubated for 1 h at room temperature in a humid chamber.
After the incubation time, the coverslip was mounted on a 3% agarose pad for imaging. Imaging was performed using the Leica DM6000 described above. Images were acquired using the 63×/1.4 numerical aperture (NA) objective and the LAS AF software (Leica Biosystems). The percentage of granule-containing embryos was counted.