Table 1.
Oligonucleotides used in quantitative real-time PCR.
Figure 1.
Supplementation of WCESP prolongs survival time of C. elegans under V. cholerae infection.
Wild-type N2 worms were supplemented with 2 mg/ml WCESP. Each lifespan experiment was repeated in three independent trials with similar results. Quantitative data and statistical analyses for the representative experiments are included in Table 2.
Figure 2.
WCESP's effects on V. cholerae.
(A) Growth of V. cholerae C 6706 in LB medium with and without 2 mg/ml WCESP. The experiment was repeated in three independent trials with similar results. Data shown here are representatives from one experiment. (B) WCESP slightly reduces colonization within C. elegans intestine. Data presented are averages from three independent trials, and error bars are standard error of the mean. P value was calculated using Student's t-test. *, p<0.05 when compared to the corresponding control. (C) qRT-PCR analysis of major V. cholerae virulence-related genes in response to 2 mg/ml WCESP (normalized to 16S rRNA). Results are the average of three independent experiments, and error bars are standard error of the mean.
Table 2.
Survival of C. elegans N2 and various mutation strains at 25°C.
Figure 3.
qRT-PCR analysis of C. elegans innate immunity genes in response to 2 mg/ml WCESP supplementation (normalized to act-1).
Results are the average of three independent experiments, and error bars are standard error of the mean. *, p<0.05 when compared to the non-treated control. **, p<0.01 when compared to the non-treated control.
Figure 4.
WCESP-mediated protection against V. cholerae is dependent on the p38 MAPK signaling pathway.
(A) pmk-1(km25) and (B) sek-1(ag1) mutant worms were supplemented with 2 mg/ml WCESP. Each lifespan experiment was repeated in at least three independent trials with similar results. Quantitative data and statistical analyses for the representative experiments are included in Table 2. (C) Western blot analysis of the phosphorylated PMK-1 levels from WCESP untreated and treated worms. Actin protein was used as the loading control.
Figure 5.
WCESP-mediated protection against V. cholerae requires HSF-1 and the IIS, but not Daf-16.
(A) daf-16(mgDf50), (B) hsf-1(sy441), (C) daf-2(e1370), and (D) age-1(hx546) mutant worms were supplemented with 2 mg/ml WCESP. Each lifespan experiment was repeated in at least three independent trials with similar results. Quantitative data and statistical analyses for the representative experiments are included in Table 2.
Figure 6.
Putative mechanisms of WCESP-mediated protection against V. cholerae and other possible bacterial pathogens in C. elegans model.
WCESP can stimulate the innate immunity by activating the p38 MAPK signaling pathway and by suppressing the IIS pathway which leads to an increase in HSF-1 activity.
Table 3.
Survival of C. elegans N2 on other pathogenic bacterial strains at 25°C.