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A A schematic diagram illustrating the de novo pyrimidine synthesis signaling pathway. B Representative images (left) and quantification results (right) of the PKA sensor worms after treatment with 100 μM <t>teriflunomide,</t> initiated from the L1 larval stage through adulthood. The scale bar indicates a length of 100 μm. C Western blot analysis of worms treated with Teriflunomide. D The quantification of PKA activities resulting from the knockdown of genes involved in the pyrimidine synthesis pathway. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM. E Western blot analysis of dhod-1 and umps-1 RNAi worms. F Supplementation of orotic acid suppressed the effect of dhod-1 RNAi on the activation of intestinal PKA but not the umps-1 RNAi. N = 3 independent experiments. The statistical significance values were determined by two-way ANOVA. Error bars denote the SEM. G Measurement of cAMP levels in C. elegans following treatment with indicated RNAi. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM.
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A A schematic diagram illustrating the de novo pyrimidine synthesis signaling pathway. B Representative images (left) and quantification results (right) of the PKA sensor worms after treatment with 100 μM teriflunomide, initiated from the L1 larval stage through adulthood. The scale bar indicates a length of 100 μm. C Western blot analysis of worms treated with Teriflunomide. D The quantification of PKA activities resulting from the knockdown of genes involved in the pyrimidine synthesis pathway. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM. E Western blot analysis of dhod-1 and umps-1 RNAi worms. F Supplementation of orotic acid suppressed the effect of dhod-1 RNAi on the activation of intestinal PKA but not the umps-1 RNAi. N = 3 independent experiments. The statistical significance values were determined by two-way ANOVA. Error bars denote the SEM. G Measurement of cAMP levels in C. elegans following treatment with indicated RNAi. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM.

Journal: Communications Biology

Article Title: Genome-wide RNAi screening in C. elegans reveals OXPHOS and pyrimidine synthesis pathways as PKA regulators

doi: 10.1038/s42003-025-08718-0

Figure Lengend Snippet: A A schematic diagram illustrating the de novo pyrimidine synthesis signaling pathway. B Representative images (left) and quantification results (right) of the PKA sensor worms after treatment with 100 μM teriflunomide, initiated from the L1 larval stage through adulthood. The scale bar indicates a length of 100 μm. C Western blot analysis of worms treated with Teriflunomide. D The quantification of PKA activities resulting from the knockdown of genes involved in the pyrimidine synthesis pathway. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM. E Western blot analysis of dhod-1 and umps-1 RNAi worms. F Supplementation of orotic acid suppressed the effect of dhod-1 RNAi on the activation of intestinal PKA but not the umps-1 RNAi. N = 3 independent experiments. The statistical significance values were determined by two-way ANOVA. Error bars denote the SEM. G Measurement of cAMP levels in C. elegans following treatment with indicated RNAi. N = 3 independent experiments. The statistical significance values were determined by one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. Error bars denote the SEM.

Article Snippet: Forskolin (FSK, T2939, purity 99.86%), Teriflunomide (T7534, purity 99.95%), and Rotenone (T2970, purity 99.88%) were purchased from TargetMol Company.

Techniques: Western Blot, Knockdown, Activation Assay

A Representative (left) and quantitative (right) Western blot results of DHODH knockdown cells treated with 500 μM orotic acid. B and C Western blot analysis on HEK293T cells treated with teriflunomide in a dose-dependent ( B ) and time-dependent ( C ) manner. N = 3 independent experiments, statistical significance determined by two-way ANOVA. Error bars represent SEM.

Journal: Communications Biology

Article Title: Genome-wide RNAi screening in C. elegans reveals OXPHOS and pyrimidine synthesis pathways as PKA regulators

doi: 10.1038/s42003-025-08718-0

Figure Lengend Snippet: A Representative (left) and quantitative (right) Western blot results of DHODH knockdown cells treated with 500 μM orotic acid. B and C Western blot analysis on HEK293T cells treated with teriflunomide in a dose-dependent ( B ) and time-dependent ( C ) manner. N = 3 independent experiments, statistical significance determined by two-way ANOVA. Error bars represent SEM.

Article Snippet: Forskolin (FSK, T2939, purity 99.86%), Teriflunomide (T7534, purity 99.95%), and Rotenone (T2970, purity 99.88%) were purchased from TargetMol Company.

Techniques: Western Blot, Knockdown