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. 2019 Jul 26;20(15):3677. doi: 10.3390/ijms20153677

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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ADAM10 retransfected in HEK293T ADAM 10/17^−/− cells is capable of shedding the IL-11R. HEK293T wild-type cells (lanes 1–3) and HEK293T ADAM10/17^−/− cells (lanes 4–6) were transfected with a N-terminally myc-tagged variant of the IL-11R and stimulated with phorbol myristate acetate (PMA) or ionomycin. Dimethyl sulfoxide (DMSO) served as control. HEK293T ADAM10/17^−/− were transfected with an N-terminally myc-tagged variant of the IL-11R and ADAM10 or ADAM17 (lanes 7–9). For both setups the supernatant was ultracentrifuged, TCA-precipitated, and analyzed by immunoblotting with different antibodies. ADAM10 flag-tagged, IL-11R myc-tagged, and ADAM17 flag-/myc-tagged were detected in lysate controls. pcDNA3.1 served as mock and actin as loading control.