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. 2000 Sep 5;192(5):729–740. doi: 10.1084/jem.192.5.729

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JEM000333.f6a.jpg — Akt regulates cytokine secretion in FcεRI-stimulated mast cells. Retroviruses coding for HA-tagged wt or K179M Akt were used to overexpress Akt proteins in stably transfected wt BMMCs. (A) Expression of HA-tagged Akt and endogenous Akt proteins was confirmed by immunoblot analysis of lysates or immune complexes precipitated with anti-HA or anti-Akt. Stable transfectants were sensitized with anti-DNP IgE and stimulated with DNP-HSA for the indicated periods of time. Cell lysates were also analyzed for Akt Ser-473 phosphorylation. Stim., stimulation; vec, vector. (B) Transfectants were sensitized with anti-DNP IgE and stimulated with the indicated concentrations of DNP-HSA for 20 h. IL-2 and TNF-α secreted into culture media were measured by ELISA. Values of secreted cytokines varied <10% from the averages among triplicate samples, and therefore SDs were not shown. Similar results were observed in another set of transfectants.

graphic file with name JEM000333.f6b.jpg — Akt regulates cytokine secretion in FcεRI-stimulated mast cells. Retroviruses coding for HA-tagged wt or K179M Akt were used to overexpress Akt proteins in stably transfected wt BMMCs. (A) Expression of HA-tagged Akt and endogenous Akt proteins was confirmed by immunoblot analysis of lysates or immune complexes precipitated with anti-HA or anti-Akt. Stable transfectants were sensitized with anti-DNP IgE and stimulated with DNP-HSA for the indicated periods of time. Cell lysates were also analyzed for Akt Ser-473 phosphorylation. Stim., stimulation; vec, vector. (B) Transfectants were sensitized with anti-DNP IgE and stimulated with the indicated concentrations of DNP-HSA for 20 h. IL-2 and TNF-α secreted into culture media were measured by ELISA. Values of secreted cytokines varied <10% from the averages among triplicate samples, and therefore SDs were not shown. Similar results were observed in another set of transfectants.