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In parasitic helminths, resistance to macrocyclic lactones (MLs) has been associated with\n                      <jats:italic>pgp<\/jats:italic>\n                      regulation and structural defects in amphids. In\n                      <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                      , the nuclear hormone receptor (NHR)-8 also influences xenobiotic tolerance by regulating\n                      <jats:italic>pgp<\/jats:italic>\n                      genes. However, the specific contribution of individual transporters and their regulation remain poorly defined. We recently demonstrated that PGP-9 specifically contributes to ivermectin (IVM) tolerance in an IVM-resistant\n                      <jats:italic>C. elegans<\/jats:italic>\n                      strain. This study aimed to explore the role of PGP-9 in drug efflux in\n                      <jats:italic>C. elegans<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We used the IVM-resistant and dye-filling defective\u00a0(Dyf)\n                      <jats:italic>C. elegans<\/jats:italic>\n                      strain IVR10 and a\n                      <jats:italic>pgp-9<\/jats:italic>\n                      IVR10 mutant to assess larval development under MLs (eprinomectin (EPR) and moxidectin (MOX)) and tunicamycin (TM). We evaluated whether the Dyf phenotype\u00a0was affected by\n                      <jats:italic>pgp-9<\/jats:italic>\n                      deletion. We investigated the role of NHR-8 in regulating\n                      <jats:italic>pgp-9<\/jats:italic>\n                      via reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and by assessing ML sensitivity in an IVR10\n                      <jats:italic>nhr-8<\/jats:italic>\n                      mutant. Additional candidate regulators of\n                      <jats:italic>pgp-9<\/jats:italic>\n                      were also tested.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      IVR10 displayed resistance to MLs and to TM, while\n                      <jats:italic>pgp-9<\/jats:italic>\n                      deletion restored full drug sensitivity despite the persistence of the Dyf phenotype. Although\n                      <jats:italic>nhr-8<\/jats:italic>\n                      deletion in IVR10 increased IVM sensitivity,\n                      <jats:italic>pgp-9<\/jats:italic>\n                      expression was not significantly altered in that strain or IVR10. Interfering RNA (RNAi) targeting\n                      <jats:italic>pgp-9<\/jats:italic>\n                      in the\n                      <jats:italic>nhr-8<\/jats:italic>\n                      mutant further increased IVM sensitivity, uncoupling PGP-9 from NHR-8 regulation. Candidate NHRs did not affect IVM tolerance in N2B.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      These results provide the first direct evidence that PGP-9 is necessary for multidrug tolerance in\n                      <jats:italic>C. elegans<\/jats:italic>\n                      , independently of amphid structural defects and NHR-8 regulation. These findings uncover a novel mechanism supporting drug resistance and highlight PGP-9 as a potential therapeutic target to improve ML treatments.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-025-07091-5","type":"journal-article","created":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T09:20:43Z","timestamp":1761816043000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["P-glycoprotein-9-mediated multidrug tolerance in Caenorhabditis elegans"],"prefix":"10.1186","volume":"18","author":[{"given":"Clara","family":"Blancfuney","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Guchen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Lespine","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e9lanie","family":"Alberich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,10,30]]},"reference":[{"key":"7091_CR1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2022.114542","volume":"239","author":"K Engle","year":"2022","unstructured":"Engle K, Kumar G. 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