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Here, we investigated by whole-cell patch-clamp measurements on HEK293 cells co-expressing ClC-Ka (tagged with GFP) and the accessory subunit barttin (tagged with m-Cherry) the effect of a natural diuretic extract from roots of\n                    <jats:italic>Dandelion<\/jats:italic>\n                    (DRE), and other compounds activating PKC, such as ATP, on ClC-Ka activity and its membrane localization. Treatment with 400\u2009\u00b5g\/ml DRE significantly inhibited Cl\n                    <jats:sup>-<\/jats:sup>\n                    currents time-dependently within several minutes. Of note, the same effect on Cl\n                    <jats:sup>-<\/jats:sup>\n                    currents was obtained upon treatment with 100\u2009\u00b5M ATP. Pretreatment of cells with either the intracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    chelator BAPTA-AM (30 \u03bcM) or the PKC inhibitor Calphostin C (100\u2009nM) reduced the inhibitory effect of DRE. Conversely, 1\u2009\u00b5M of phorbol meristate acetate (PMA), a specific PKC activator, mimicked the inhibitory effect of DRE on ClC-Ka. Finally, we found that pretreatment with 30\u2009\u00b5M Heclin, an E3 ubiquitin ligase inhibitor, did not revert DRE-induced Cl\n                    <jats:sup>-<\/jats:sup>\n                    current inhibition. In agreement with this, live-cell confocal analysis showed that DRE treatment did not induce ClC-Ka internalization. In conclusion, we demonstrate for the first time that the activity of ClC-Ka in renal cells could be significantly inhibited by the activation of PKC elicited by classical maneuvers, such as activation of purinergic receptors, or by exposure to herbal extracts that activates a PKC-dependent pathway. Overall, we provide both new information regarding the regulation of ClC-Ka and a proof-of-concept study for the use of DRE as new diuretic.\n                  <\/jats:p>","DOI":"10.1038\/s41598-020-67219-8","type":"journal-article","created":{"date-parts":[[2020,6,24]],"date-time":"2020-06-24T06:02:52Z","timestamp":1592978572000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Role of PKC in the Regulation of the Human Kidney Chloride Channel ClC-Ka"],"prefix":"10.1038","volume":"10","author":[{"given":"Andrea","family":"Gerbino","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberta","family":"De Zio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniela","family":"Russo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luigi","family":"Milella","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Serena","family":"Milano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giuseppe","family":"Procino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Pusch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria","family":"Svelto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monica","family":"Carmosino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,6,24]]},"reference":[{"key":"67219_CR1","doi-asserted-by":"publisher","first-page":"1493","DOI":"10.1152\/physrev.00047.2017","volume":"98","author":"TJ Jentsch","year":"2018","unstructured":"Jentsch, T. 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