F55G11.8 : Orthosiphon stamineus aqueous extract : clec-52 : clec-60 : clec-67 : cpr-2 : dod-22 : dod-24 : irg-5 : lys-7 : sod-3 : spp-1

"We measured mRNA levels of 11 immune effectors in worms grown in the presence and absence of UE-12 by qRT-PCR. Interestingly, the expression of none of these genes was significantly altered by UE-12 (supplementary material Fig. S5), implying that UE-12 does not affect the basal expression of these genes."

C17H12.8 : F55G11.8 : Staphylococcus aureus infection : clec-67 : dod-24 : irg-5 : sysm-1

"we performed transcriptome analysis on a selected panel of PMK-1 and DAF- 16-regulated genes using quantitative real-time PCR. Six of the eight PMK-1-regulated genes were downregulated (~2-5-fold) following 48 hours infection by S. aureus of worms that were not treated with UE-12 (Fig. 5A). On the other hand, in infected worms treated with 200 mg/mL UE-12, five of these 6 genes - clec-67, C17H12.8, dod-24, F35E12.5 and F55G11.8 - demonstrated a 2.8-4-fold induction in expression upon supplementation with UE-12 (p<0.05), implying that UE-12 promoted the host antimicrobial response. UE-12 also significantly reduced the suppression of T24B8.5 from ~5-fold to ~2-fold (p<0.05) (Fig. 5A)."

C17H12.8 : F55G11.8 : Orthosiphon stamineus aqueous extract : clec-67 : dod-24 : irg-5 : sysm-1

"we performed transcriptome analysis on a selected panel of PMK-1 and DAF- 16-regulated genes using quantitative real-time PCR. Six of the eight PMK-1-regulated genes were downregulated (~2-5-fold) following 48 hours infection by S. aureus of worms that were not treated with UE-12 (Fig. 5A). On the other hand, in infected worms treated with 200 mg/mL UE-12, five of these 6 genes - clec-67, C17H12.8, dod-24, F35E12.5 and F55G11.8 - demonstrated a 2.8-4-fold induction in expression upon supplementation with UE-12 (p<0.05), implying that UE-12 promoted the host antimicrobial response. UE-12 also significantly reduced the suppression of T24B8.5 from ~5-fold to ~2-fold (p<0.05) (Fig. 5A)."

Eurycoma longifolia root water extract : lys-7

"we tested the effect of these candidates on the expression of a host immune effector using transgenic worms carrying a transcriptional GFP reporter for the lys-7 gene... The transgenic animals were infected by S. aureus in the presence and absence of antiinfective candidates and fluorescence intensity was visualized and compared at 24 and 48 hours post-infection (hpi). As shown in Fig. 1A and Fig. 1B, at 48 hpi, nematodes infected by S. aureus in the absence of anti-infective candidates showed a significant reduction in the overall GFP intensity as compared to the uninfected population fed on the normal food source, Escherichia coli OP50. The uninfected worms showed a consistently strong green fluorescence along the intestine throughout the observation period (Fig. 1A) whereas S. aureus-infected worms exhibited weak fluorescence except at the posterior part of the intestine (Fig. 1B). Interestingly, among the anti-infective candidates tested, the UE-12 extract was able to prevent suppression of GFP expression by S. aureus (Fig. 1C). By contrast, UE-01-4, UE-03-6, UE-11 and UC-10 failed to prevent the suppression of GFP expression by S. aureus (supplementary material Fig. S1)."

dod-22 : lys-7 : sma-6

"we also tested the expression of these genes in UE-12-treated and untreated sma-6 (wk7) mutant worms infected by S. aureus. Similar to the profile observed in wild-type N2 worms, in a sma-6 mutant background, the suppression of lys-7 and dod-22 by S. aureus was significantly induced in the presence of UE-12 extract (supplementary material Fig. S4)"

R13H8.3 : R13H8.5 : R13H8.6 : daf-16 : dod-22 : mtl-1 : sod-3 : spp-1

"In the absence of UE-12, spp-1 and dod-22 were downregulated (2-3-fold), mtl-1 was upregulated (~2.5-fold) whilst sod-3 was unaffected (Fig. 5C). In infected worms treated with UE-12, all four genes were significantly induced by 3.3-7.5-fold (p<0.05) (Fig. 5C), indicating that UE-12 was able to boost the expression of DAF-16-regulated genes. To confirm that the modulation of these genes require the presence of DAF-16, we profiled the expression of all four DAF-16 targets as well as lys-7 (Fig. 1H) in the daf-16 (mu86) mutant background. No significant difference was observed in the expression of these genes in both treated and untreated daf-16 mutant animals (Fig. 5D)."

Staphylococcus aureus infection : mtl-1

"In the absence of UE-12, spp-1 and dod-22 were downregulated (2-3-fold), mtl-1 was upregulated (~2.5-fold) whilst sod-3 was unaffected (Fig. 5C). In infected worms treated with UE-12, all four genes were significantly induced by 3.3-7.5-fold (p<0.05) (Fig. 5C), indicating that UE-12 was able to boost the expression of DAF-16-regulated genes. To confirm that the modulation of these genes require the presence of DAF-16, we profiled the expression of all four DAF-16 targets as well as lys-7 (Fig. 1H) in the daf-16 (mu86) mutant background. No significant difference was observed in the expression of these genes in both treated and untreated daf-16 mutant animals (Fig. 5D)."

Orthosiphon stamineus aqueous extract : clec-52 : clec-60 : cpr-2 : cyp-37B1

"We found that during the late stage of infection (48 hpi), expression of clec-52, clec-60, ilys-3 and cyp-37B1 was downregulated, an effect that was reversed when infected worms were exposed to UE-12 (Fig. 5E). Specifically, the expression of clec-52, clec-60 and cyp-37B1 was induced significantly (p,0.05). Moreover, cpr-2 demonstrated a further induction (p,0.05). At this late time point, the expression of ilys-3, F53A9.8 and fmo-2 remained unchanged in treated and untreated animals (Fig. 5E). The fold difference for genes significantly modulated by UE-12 is shown in Table 1."

Nypa fruticans root butanol extract : lys-7

"we tested the effect of these candidates on the expression of a host immune effector using transgenic worms carrying a transcriptional GFP reporter for the lys-7 gene... The transgenic animals were infected by S. aureus in the presence and absence of antiinfective candidates and fluorescence intensity was visualized and compared at 24 and 48 hours post-infection (hpi). As shown in Fig. 1A and Fig. 1B, at 48 hpi, nematodes infected by S. aureus in the absence of anti-infective candidates showed a significant reduction in the overall GFP intensity as compared to the uninfected population fed on the normal food source, Escherichia coli OP50. The uninfected worms showed a consistently strong green fluorescence along the intestine throughout the observation period (Fig. 1A) whereas S. aureus-infected worms exhibited weak fluorescence except at the posterior part of the intestine (Fig. 1B). Interestingly, among the anti-infective candidates tested, the UE-12 extract was able to prevent suppression of GFP expression by S. aureus (Fig. 1C). By contrast, UE-01-4, UE-03-6, UE-11 and UC-10 failed to prevent the suppression of GFP expression by S. aureus (supplementary material Fig. S1)."

Staphylococcus aureus infection : lys-7

"we tested the effect of these candidates on the expression of a host immune effector using transgenic worms carrying a transcriptional GFP reporter for the lys-7 gene... The transgenic animals were infected by S. aureus in the presence and absence of antiinfective candidates and fluorescence intensity was visualized and compared at 24 and 48 hours post-infection (hpi). As shown in Fig. 1A and Fig. 1B, at 48 hpi, nematodes infected by S. aureus in the absence of anti-infective candidates showed a significant reduction in the overall GFP intensity as compared to the uninfected population fed on the normal food source, Escherichia coli OP50. The uninfected worms showed a consistently strong green fluorescence along the intestine throughout the observation period (Fig. 1A) whereas S. aureus-infected worms exhibited weak fluorescence except at the posterior part of the intestine (Fig. 1B). Interestingly, among the anti-infective candidates tested, the UE-12 extract was able to prevent suppression of GFP expression by S. aureus (Fig. 1C). By contrast, UE-01-4, UE-03-6, UE-11 and UC-10 failed to prevent the suppression of GFP expression by S. aureus (supplementary material Fig. S1)."