NPR-15 loss-of-function enhanced pathogen resistance and inhibited avoidance behavior.

A. Wild type (WT), npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals were exposed to P. aeruginosa partial lawn and scored for survival.

B. Colonization of WT, npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals by P. aeruginosa-GFP after 24 hours at 25°C. Scale bar, 200 μm.

C. Colony-forming units per animal [WT, npr-15(tm12539), npr-15(tm12539);Pnp-15::npr-15] grown on P. aeruginosa-GFP for 24 hours at 25°C. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.001 and NS = Not Significant.

D. WT, npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals were exposed to S. enterica partial lawn and scored for survival.

E. WT, npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals were exposed to E faecalis partial lawn and scored for survival.

F. WT, npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals were exposed to S. aureus partial lawn and scored for survival.

G. WT, npr-15(tm12539), and npr-15(tm12539);Pnp-15::npr-15 animals were exposed to live E. coli and scored for survival.

H. WT and npr-15(tm12539) animals were exposed to UV-killed E. coli and scored for survival.

I. Schematics of avoidance behavior assay on S. aureus.

J. Lawn occupancy of WT C. elegans and npr-15(tm12539), and npr-15(tm1250);Pnpr-15::npr-15 animals on a partial lawn of S. aureus at 24Lhours. Bars represent means while error bars indicate SD; **p < 0.001 and NS = Not Significant.

NPR-15 inhibits the expression of immune and aversion-related genes/pathways.

A. Volcano plot of upregulated and downregulated genes in npr-15(tm12539) vs. WT animals. Red and blue dots represent significant upregulated and downregulated genes respectively, while the grey dots represent not significant genes.

B. Gene ontology analysis of upregulated genes in npr-15(tm12539) vs. WT animals. The result was filtered based on significantly enriched terms, with a Q value < 0.1.

C. Representation factors of immune pathways for the upregulated immune genes in npr-15(tm12539) vs. WT animals.

D. Venn diagram showing the upregulated immune genes in each pathway in npr-15(tm12539) vs. WT animals.

E. qRT-PCR analysis of ELT-2-depenent immune gene expression in WT and npr-15(tm12539) animals. Bars represent means while error bars indicate SD of three independent experiments; *p < 0.05, **p < 0.001, and ***p < 0.0001.

F. qRT-PCR analysis of HLH-30-depenent immune gene expression in WT and npr-15(tm12539) animals. Bars represent means while error bars indicate SD of three independent experiments; *p < 0.05, **p < 0.001, and ***p < 0.0001.

NPR-15 loss-of-function enhances immunity via ELT-2 and HLH-30 when exposed to S. aureus.

A. WT and npr-15(tm12539) animals fed with elt-2 RNAi were exposed to S. aureus full lawn and scored for survival. EV, empty vector RNAi control.

B. WT and npr-15(tm12539) animals fed with hlh-30 RNAi were exposed to S. aureus full lawn and scored for survival. EV, empty vector RNAi control.

C. WT and npr-15(tm12539) animals fed with hlh-30 and elt-2 RNAi were exposed to S. aureus full lawn and scored for survival. EV, empty vector RNAi control.

D. WT and npr-15(tm12539) animals fed with pmk-1 RNAi and animals were exposed to S. aureus full lawn and scored for survival. EV, empty vector RNAi control.

E. WT and npr-15(tm12539) animals fed with daf-16 RNAi were exposed to S. aureus full lawn and scored for survival. EV, empty vector RNAi control.

NPR-15 suppresses molecular immune response via sensory neurons, ASJ.

A. Neuronal connectome of NPR-15-expressing neurons.

B. WT, npr-15(tm12539), and npr-15(tm12539);Prab-3::npr-15 animals were exposed to S. aureus full lawn and scored for survival.

C. WT, npr-15(tm12539), ASJ(-), and ASJ(-);npr-15(tm12539) animals were exposed to S. aureus full lawn and scored for survival.

D. WT, npr-15(tm12539), ASG(-), and ASG(-);npr-15(tm12539) animals were exposed to S aureus full lawn and scored for survival.

E. WT, npr-15(tm12539), ASE(-), and ASE(-);npr-15(tm12539) animals were exposed to S. aureus full lawn and scored for survival.

F. WT, npr-15(tm12539), npr-15(tm12539);Ptrx-1::npr-15 animals were exposed to S. aureus full lawn and scored for survival.

G. qRT-PCR analysis of ELT-2 and HLH-30-depenent immune gene expression in WT and ASJ(-) animals. Bars represent means while error bars indicate SD of three independent experiments; *p < 0.05, **p < 0.001 and ***p < 0.0001.

NPR-15 loss-of-function inhibits pathogen avoidance behavior in an ASJ-dependent manner.

A. WT, npr-15(tm12539), and npr-15(tm12539);Prab-3::npr-15 animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

B. WT, npr-15(tm12539), ASJ(-), and ASJ(-);npr-15(tm12539) animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

C. WT, npr-15(tm12539), and npr-15(tm12539);Ptrx-1::npr-15 animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

D. WT, npr-15(tm12539), ASE(-), and ASE(-);npr-15(tm12539) animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

E. WT, npr-15(tm12539), ASI(-), and ASI(-);npr-15(tm12539) animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; **p < 0.001, ***p < 0.0001, and NS = Not Significant.

F. WT, npr-15(tm12539), and AFD(-), and AFD(-);npr-15(tm12539) animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

G. WT, npr-15(tm12539), and AWC(-), and AWC(-);npr-14(tm12539) animals were exposed to partial lawn of S. aureus and scored for lawn occupancy. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

H. WT and npr-15(tm12539) fed with gon-2 RNAi and were exposed to the partial lawn of S. aureus and scored for lawn occupancy. EV, empty vector RNAi control. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001.

I. WT and npr-15(tm12539) fed with gtl-2 RNAi and were exposed to the partial lawn of S. aureus and scored for lawn occupancy. EV, empty vector RNAi control. Bars represent means while error bars indicate the SD of three independent experiments; **p < 0.001, ***p < 0.0001, and NS = Not Significant.

J. WT, npr-15(tm12539), RNAi intestine-specific strain MGH171, MGH171; npr-15(tm12539) fed with gon-2 RNAi and were exposed to the partial lawn of S. aureus and scored for lawn occupancy. EV, empty vector RNAi control. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001 and NS = Not Significant.

K. WT, npr-15(tm12539), ASJ(-), ASJ(-);npr-15(tm12539) fed with gon-2 RNAi and were exposed to the partial lawn of S. aureus and scored for lawn occupancy. EV, empty vector RNAi control. Bars represent means while error bars indicate the SD of three independent experiments; ***p < 0.0001.

GPCR/NPR-15 suppressed immune response and enhanced avoidance behavior via sensory neurons, ASJ.

The immune response control is dependent on ELT-2 and HLH-30 transcription factors. While NPR-15 controls avoidance behavior to S. aureus via intestinal-expressed TRPM channel, GON-2.