Fig 1.
FSHR-1 is required for the defense against diverse pathogens.
Representative wild-type (A) and fshr-1(ok778) mutant (B) worms fed GFP-PA14 for 18 hours. Scale bars = 100 μm. (C) Quantification of the sum of the intestinal fluorescence intensity for at least 10 representative individual worms per condition. (D) Mean survival time of wild-type and fshr-1(ok778) mutant worms exposed to different bacterial food sources. * P<0.05, **P<0.01, *** P<0.0001 in unpaired t-test. Error bars are SEM. (E) Functional annotation of genes whose mean expression from three microarray replicates in wild-type worms infected with PA14 for 4 hours was at least 2-fold higher than their mean expression in fshr-1(ok778) mutants infected with PA14 for 4 hours, with P<0.05. Genes known to be involved both in infection response and stress response were grouped with stress response.
Fig 2.
FSHR-1 is required for the survival of heavy metal and oxidative stress.
Wild-type, fshr-1(ok778) mutants, and mutants rescued with fshr-1(+) in the intestine were measured for their ability to survive different types of stress. Worms were subjected to 100 μM cadmium (A, B), 5 mM paraquat (C, D), or 37°C (E, F). Representative survival curves for approximately 80–100 worms for each condition are shown in (A, C, and E) with standard deviation error bars. The mean survival time for each genotype was calculated for at least three independent experiments (B, D, F). Error bars are SEM. One-way ANOVA and Tukey HSD test, *P<0.05, **P<0.01.
Fig 3.
FSHR-1 is required for the induction of the oxidative stress response by PA14.
ldIs3[gcs-1::gfp] (A, B) or fshr-1(ok778); ldIs3[gcs-1::gfp] (C, D) worms were exposed to OP50 (A, C) or PA14 (B, D) for 5 hours. Representative worms show that in a wild-type background, gcs-1::gfp is induced by pathogen exposure; this induction depends on fshr-1. Scale bars = 100 μm. (E) Quantification of the mean fluorescence intensity of the intestine for each condition (see Methods). Error bars are SEM. One-way ANOVA and Tukey HSD test, **P<0.01.
Fig 4.
FSHR-1 promotes the avoidance of pathogenic PA14.
The occupancy of wild-type, fshr-1(ok778), and ok778; agIs35[gut::fshr-1(+)] worms on a lawn of relatively non-pathogenic E. coli OP50 (A, B) or pathogenic P. aeruginosa PA14 (C, D) was measured <1 hour (A, C) and 9 hours (B, D) after transferring worms to the lawn. Error bars are SEM. One-way ANOVA and Tukey HSD test, *P<0.05, **P<0.01.
Fig 5.
Pathogen avoidance contributes to the pathogen sensitivity of fshr-1 mutants.
The survival of worms was measured when exposed to a standard small lawn of PA14 or a big lawn that covered the entire surface of the agar. (A) Representative killing assay. (B) Mean survival time was measured and compared using one-way ANOVA and Tukey HSD test, **P<0.01. Error bars are standard deviation (A) or SEM (B).