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. 2016 Sep 1;9(9):1051–1061. doi: 10.1242/dmm.024745

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© 2016. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 4. — Programming of macrophages with ADPKD-CM promotes IL-10-dependent STAT3 activation. (A,B) RAW macrophages were incubated with ADPKD-CM for the time indicated before lysis and immunoblot analyses using anti-phospho-STAT3 (pSTAT3) and anti-STAT3 (STAT3) antibodies. (A) Immunoblot of cell samples collected at 0-60 min. (B) Immunoblot of cell samples collected at 1-24 h. NS, non-specific bands, which serve as a control for protein loading. Similar results were observed in three independent experiments using CM generated with cyst cells from different ADPKD individuals. (C) RAW macrophages were incubated for 18 h with either mouse IL-10 (10 ng/ml), ADPKD-CM or medium alone, as indicated, in the presence of an anti-IL-10 neutralizing antibody (αIL10) or equal concentrations of a control antibody (IgG) before cell lysis. Total cell lysates were subjected to immunoblot analyses using an anti-phospho-STAT3 antibody, as well as anti-tubulin antibodies for a control. Similar results were observed in three independent experiments using CM generated with cyst cells from different ADPKD individuals.