Extended Data Fig. 2. Pathogen exposure induces pheromone response of AWA neurons.

a - d, Traces of GCaMP6 signals in AWA neurons of wild-type adult hermaphrodites in response to pheromones at different concentrations when exposed to OP50 or PA14 (a,c) and quantitation of average GCaMP6 signals in AWA during pheromone stimulation in the previous panel (b,d). Phe, pheromone mixture of ascr#2,3,5. Lines in traces, mean. Shades, s.e.m.. F_baseline, average GCaMP6 signals in the first 30 seconds. Box plot, median, 1^st and 3^rd quartiles; whiskers, minimum and maximum. Numbers in parenthesis, number of individual worms. Dots, average GCaMP6 signals of individual worms during pheromone stimulation. P values are derived from two-tailed Mann-Whitney test, asterisks indicate significant difference, **** P < 0.0001, ** P < 0.01.

e, Paradigm of a long-term exposure to bacterial pathogen PA14.

f, Avoidance of ascaroside pheromones in wild-type adult hermaphrodites exposed to OP50 or PA14 in a long-term exposure paradigm as shown in (e). Pheromones, mixture of ascr#2,3,5 (10 nM each at equilibrium). Positive avoidance index, avoidance. Numbers in parenthesis, number of individual assays. Box plot, median, 1^st and 3^rd quartiles; whiskers, minimum and maximum. Dots, avoidance indexes of individual assays. P values are derived from two-tailed unpaired t test, asterisks indicate significant difference, ** P < 0.01.

g, h, Traces of GCaMP6 signals in AWA neurons of wild-type adult hermaphrodites in response to ascaroside pheromones when exposed to OP50 or PA14 in the long-term exposure paradigm (g) and quantitation of average GCaMP6 signals in AWA during pheromone stimulation (h). Phe, pheromones, individual or mixture of ascr#2,3,5 (1 μM each). Lines in traces, mean. Shades, s.e.m.. F_baseline, average GCaMP6 signals in the first 30 seconds. Box plot, median, 1^st and 3rd quartiles; whiskers, minimum and maximum. Numbers in parenthesis, number of individual worms. Dots, average GCaMP6 signals of individual worms during pheromone stimulation. P values are derived from One-way ANOVA with Dunnett’s multiple comparisons test, asterisks indicate significant difference, **** P < 0.0001, ** P < 0.01, ns, not significant.

i – l, Traces of GCaMP6 signals in AWA neurons of unc-13(e51) (i) or unc-31(e928) (k) mutant adult hermaphrodites in response to pheromones when exposed to OP50 or PA14 and quantitation of average GCaMP6 signals during pheromone stimulation (j,l) in the previous panel. Phe, Pheromone mixture of ascr#2,3,5 (1 μM each). Lines in traces, mean. Shades, s.e.m.. F_baseline, average GCaMP6 signals in the first 30 seconds. Box plot, median, 1^st and 3rd quartiles; whiskers, minimum and maximum. Numbers in parenthesis, number of individual worms. Dots, average GCaMP6 signals of individual worms during pheromone stimulation. P values are derived from two-tailed unpaired t test, asterisks indicate significant difference, **** P < 0.0001, ** P < 0.01. P values are shown in Source data.