Table 1.

Time course of immobility response and effect of octanol

General protocol
NGM (5X peptone) plate plus food, 15° C
↓
NGM (5X peptone) plate plus food, 25° C overnight (16 – 18 h)
↓
NGM plate plus food (± DMSO + solvent or drug), 25°C for 90 min
↓
NGM plate minus food (± DMSO + solvent or drug), 25°C for the observation period

	Moving/total
	1 h	3 h	24 h	48 h	72 h	96 h
daf-2(e1391) DMSO, no food	5/38
daf-2(e1391) DMSO, no food + octanol challenge	30/34
N2 DMSO, no food	25/26	19/20	8/8	6/7	3/3	0a
daf-2(e1391) DMSO, no food	3/22	4/20	0/20	1/22	1/22	0/21
unc-64(e246) DMSO, no food	4/24	2/25	3/25	1/25	1/22	0/22

Animals (25–40 per plate) were transferred to bacteria-free NGM plates containing DMSO, as shown at the top of the table. For the octanol experiments, we dipped a bristle from a horsehair paintbrush in octanol, held this in front of the nose of the immobile animals, and tallied whether they backed away as expected. For the time course experiments, we periodically assessed spontaneous movement of animals over 96 h. At all time points, the animals still escaped normally in response to tail touch. The steady decline in numbers in the control (N2) group reflected animals that attempted to leave the no-food environment, crawled up the side of the plate, and desiccated off the agar. This represents a typical response of Caenorhabditis elegans strains with a normal drive for food seeking. NGM, nematode growth medium.

^aNo animals were left on the plate at this time point.