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. 2019 Feb 4;10:59. doi: 10.3389/fphar.2019.00059

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Copyright © 2019 Palikhe, Ohashi, Sakamoto, Hattori, Kawakami, Andoh, Yamazaki and Hattori.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of GRK2 inhibitor treatment on iNOS expression, STAT1/3 activation, and IRF1 activation in microglial cells. GRK2 inhibitor at a concentration of 10 μM was added at 30 min before challenge with 100 ng/ml LPS or 10 ng/ml IFN-β. (A) Expression of iNOS protein before and 15 h after LPS application. (B) STAT1 and STAT3 phosphorylation before and 6 h after LPS application. (C) STAT1 and STAT3 phosphorylation before and 15–30 min after IFN-β supplementation. (D) Expression of IRF1 expression before and 1 h after LPS application. (E) Cytoplasmic (C) and nuclear (N) fractions were isolated, and then changes in IRF1 levels in each fraction before and 1 h after LPS application was tracked by Western blot analysis. GAPDH served as loading control and lamin B was used as a nuclear marker. Shown are representative Western blots from three independent experiments in which the same results were obtained.