Journal: iScience

Article Title: Adenosine deaminase and deoxyadenosine regulate intracellular immune response in C. elegans

doi: 10.1016/j.isci.2025.111950

Figure Lengend Snippet: Resistance against intracellular pathogens is increased by adah-1 knockdown or deoxyadenosine treatment (A) Quantification of Orsay virus pathogen load at 18 hpi of L4 wild-type N2 animals after being treated with empty vector control RNAi, adah-1 , or pnp-1 RNAi, and a pnp-1 mutant ( jy121 ) treated with control RNAi. Infected animals were stained with an FISH probe to label Orsay virus RNA1 and RNA2, and each dot in the graph shows fluorescence for an individual animal across three independent replicates. WT, control n = 40; WT, adah-1 (RNAi) n = 42; WT, pnp-1 (RNAi) n = 40; pnp-1(−/−) control n = 36. Images for quantification were taken with a Zeiss AxioImager; see Figure S4 A. (B) Quantification of N. parisii pathogen load at 3 hpi in wild-type animals treated with control, adah-1 , or pnp-1 RNAi and a pnp-1 ( jy121 ) mutant treated with control RNAi. Each dot represents the number of sporoplasms for an individual animal; 300 animals per condition were quantified across three independent replicates, with the average sporoplasm number listed at the top. (C) Quantification of Orsay virus pathogen load 18 hpi determined by mean FISH probe fluorescence in adult rde-1(ne219) mutant animals (which are RNAi-defective and thus are more susceptible to infection than wild-type animals) that have been treated with 30 mM deoxyadenosine (dAdo) for 42 h. Control n = 155; dAdo n = 160. Images for quantification were taken with an ImageXpress plate reader; see Figure S6 A. (D) Quantification of N. parisii pathogen load 30 hpi determined by mean FISH probe fluorescence of adult wild-type N2 animals that have been treated with 30 mM dAdo for 54 h. Three biological replicates were performed. Control n = 164, dAdo n = 151. (A–D) Kruskal-Wallis test was used to calculate p values; ∗∗∗∗ p < 0.0001, ∗ p < 0.05. (E) Quantification of E. intestinalis pathogen load at 27 hpi after 48 h treatment of human endothelial cells with the indicated concentrations of deoxyadenosine. At least 118 vacuoles were quantified per condition, across three biological replicates. (F) Quantification of E. intestinalis pathogen load at 27 hpi after 48 h treatment of human endothelial cells with 0.5 mM deoxyadenosine and anti-IFNalphaR1 antibody. At least 275 vacuoles were quantified per condition, across three biological replicates. Pathogen load was determined by measuring FISH probe fluorescence area.

Article Snippet: E. intestinalis 16S rRNA FISH probe Quasar 570 , LGC Biosearch Technologies , Antao et al. .

Techniques: Knockdown, Virus, Plasmid Preparation, Control, Mutagenesis, Infection, Staining, Fluorescence

Journal: iScience

Article Title: Adenosine deaminase and deoxyadenosine regulate intracellular immune response in C. elegans

doi: 10.1016/j.isci.2025.111950

Figure Lengend Snippet:

Article Snippet: E. intestinalis 16S rRNA FISH probe Quasar 570 , LGC Biosearch Technologies , Antao et al. .

Techniques: Purification, Virus, Recombinant, Cell Culture, Control, Plasmid Preparation, Software, Microscopy