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. 2018 Nov 16;8:16931. doi: 10.1038/s41598-018-35184-y

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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APC binds PKA and responses to cAMP. Panel (A) APC binds PKA cat in RAW 264.7 cells. Immunoprecipitations were executed with control rabbit IgG, rabbit IgG against APC, or rabbit IgG against PKA cat α. The subsequent immunoblots were probed with antibodies against APC or PKA cat α. Panel (B) Elevated cAMP in macrophages leads to the release of PKA cat α from APC. Immunoprecipitations were performed using protein extracts taken from RAW 264.7 cells exposed 1 mM 6-MB-cAMP or 10 nM ET. Immunoprecipitations were carried out with control rabbit IgG or rabbit IgG against APC. The immunoblots were probed with antibodies recognizing PKA cat α. Panel (C) APC binds PKA RII in RAW 264.7 cells and human monocytes. Immunoprecipitations were performed using protein extracts from RAW 264.7 cells or primary human macrophages using control IgG or IgG against APC, and immunoblots were probed with antibodies recognizing PKA RII. Panel (D) Domain layout of PKA RII highlighting the docking and dimerization (D/D) domain that is deleted from PKA RII. Panel (E) RAW 264.7 cells were transduced with a replication incompetent retrovirus containing RII HA-tagged or ΔD/D RII HA-tagged. Immunoprecipitations were then performed with control rabbit IgG or rabbit IgG against APC. The immunoblots were probed with antibodies recognizing the HA-tag.