a, b, n, Avoidance of ascaroside pheromones when exposed to OP50 or PA14 (a,n) and modulation of pheromone avoidance by PA14 exposure (a,b) in wild-type and str-44 mutant hermaphrodites (a, b) or in transgenic hermaphrodites expressing wild-type str-44 DNA using str-44 promoter or AWA-selective promoter gpa-4delta6p in str-44 mutant background (b) or in transgenic hermaphrodites expressing a str-44 cDNA in ASH in wild type (n). Pheromones, mixture of ascr#2,3,5 (10 nM each at equilibrium). Positive avoidance index, avoidance. Positive modulation index, suppression of avoidance by PA14.
c - h, j - m, Traces of GCaMP6 signals in AWA neurons in response to ascaroside pheromones in wild-type and str-44 mutant hermaphrodites and in transgenic hermaphrodites expressing a wild-type str-44 DNA using a str-44 promoter or a wild-type str-44 cDNA using an AWA-specific promotor in str-44 mutant background exposed to OP50 or PA14 (c,e,g), or traces of GCaMP6 signals in ASH neurons in response to ascaroside pheromones or buffer control in transgenic hermaphrodites expressing a wild-type str-44 cDNA in ASH and in control hermaphrodites when exposed to OP50 (j,l), and quantitation of average GCaMP6 signals during pheromone stimulation (d,f,h,k) or during the second buffer stimulation (m) in the previous panel. Phe, pheromone mixture of ascr#2,3,5 (1 μM each). Lines in traces, mean. Shades, s.e.m.. Fbaseline, average GCaMP6 signals in the first 30 seconds.
i, Sample images for str-44p::str-44::gfp translational reporter expression in adult hermaphrodites exposed to OP50 or PA14. Dashed lines indicate enlarged views. GFP signals in AWA cilium are shown. Lines outline worm bodies. Scale bar, 20 μm. A, anterior. D, dorsal.
Box plot, median, 1st and 3rd quartiles; whiskers, minimum and maximum. Numbers in parenthesis, number of individual assays (a,b,n) or individual worms (d,f,h,k,m). Dots, avoidance indexes or modulation indexes of individual assays (a,b,n) or average GCaMP6 signals of individual worms during either pheromone stimulation (d,f,h,k) or the second 30-s buffer stimulation (m). P values are derived from Two-way ANOVA with Tukey’s multiple comparisons test [(avoidance index in a),d,n] or two-tailed unpaired t test [(modulation index in a),k] or Two-way ANOVA with Bonferroni’s multiple comparisons test (b,f,h) or two-tailed Mann-Whitney test (m), asterisks indicate significant difference, **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns, not significant. P values are shown in Source data.