Fig. 3 ERK5 inhibition suppresses M2 macrophage polarization, alters secreted cytokine profiles, and modifies the actin cytoskeleton. ERK5 inhibition suppresses.

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Fig. 3 ERK5 inhibition suppresses M2 macrophage polarization, alters secreted cytokine profiles, and modifies the actin cytoskeleton. ERK5 inhibition suppresses M2 macrophage polarization, alters secreted cytokine profiles, and modifies the actin cytoskeleton. (A) (i) Pharmacological inhibition of ERK5 with XMD17-109 does not alter expression of the M1 macrophage marker CD86 in J774 macrophages at rest or under M1 (LPS and IFN-γ) or M2 (IL-4) stimulating conditions. (ii) However, IL-4–driven M2 polarization is strongly reduced in the presence of XMD17-109. Significance was tested using two-way analysis of variance (ANOVA) on data from three independent replicates. Error bars show the SEM. (B) XMD17-109 inhibition of ERK5 activity in J774 macrophages increases proinflammatory IL-12p40 secretion (i) while reducing the secretion of the M2 marker Ym1 (ii). Significance was tested using a ratio-paired t test. (C) XMD17-109 inhibition of ERK5 activity in human PBMC-derived macrophages increases proinflammatory TNFα secretion (i) while reducing the secretion of the anti-inflammatory cytokine IL-10 (ii). Significance was tested using a ratio-paired t test. (D) J774 macrophages treated with XMD17-109 show a reduction in actin ruffles (revealed using Alexa 488–phalloidin; i). (ii) Ruffle disruption by XMD17-109 results in increased numbers of smaller actin-rich fragments, quantified in binary images (n = 33 images from n = 3 replicates, two-tailed t test; error bars are SEM). Images were captured using identical illumination, exposure, and postacquisition processing settings. Andrew S. Gilbert et al. Sci Adv 2017;3:e1700898 Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).