*2.4. Locomotive Behaviors Tests*

Wild-type N2 *C. elegans* were washed after exposure via dietary transfer and subjected to locomotive behavior tests. Locomotive behaviors, including body bending and head thrashing frequencies, were studied by modifying the methods used in a previous study [36]. The frequencies of body bending during 20 s and head thrashing during 1 min were manually measured and calculated. The tests were conducted at least three times independently, and 20 worms were scored per treatment in each replicate.

### *2.5. GABAergic Neuron Toxicity Assay*

Transgenic EG1285 strain *C. elegans* were washed after exposure through dietary transfer. The morphology of green fluorescent protein (GFP)-labeled GABAergic neurons was characterized using an epifluorescence microscope. Fluorescent images of the worms were captured for the determination of neuron cell and gap numbers in GABAergic D-type motor neurons. The abnormality of GABAergic motor neurons (%) was defined as the total counts of GABAergic motor neuron cell loss and gap numbers divided by the total GABAergic motor neuron cells and the connections between the neuron cells in normal conditions. The tests were conducted at least three times independently, and 20 worms were scored per treatment in each replicate.
