Fig. 4

RCT001 reverses the polarization of M2-like macrophages polarization. Purified monocytes from healthy human donors (n = 5) were differentiated into M0-like macrophages (5 days with CSF1) and then polarized into M0-like macrophages (CSF1), or M2-like macrophages (IL4) or M1-like macrophages (LPS) for 48 h. The already polarized macrophages then were treated with RCT001 (2.5 µM) in the presence of the respective cytokines (CSF1 for M0, IL4 for M2 and LPS for M1 macrophages) for 48 h. MFI: Mean Fluorescence Intensity a-c M0 and M2-like macrophages were treated with RCT001 for 48 h. a Specific M2 macrophage membrane markers (CD206, CD209, CD200R, CD163) were examined by flow cytometry. b Specific M2 macrophage mRNA markers (TIMP3, CCL13, CCL14, CCL17, CCL18, CCL22, CCL23, CCL24) were evaluated by qPCR. c. Specific cytokines secreted by M2 macrophages (CCL13, CCL22, CCL24) were determined by ELISA. d-f M0 and M1-like macrophages were treated with RCT001 for 48 h. d Specific M1 macrophage membrane markers (CD80, CD86) were evaluated by flow cytometry. e Specific M1 macrophages mRNA markers (IL8, CXCL11, CCL20, IL-1a) were evaluated by qPCR. f Specific cytokines secreted by M1 macrophages (IL8, CXCL11, CCL20, IL-1a) were determined by ELISA. Results are presented as the mean of five independent experiments ± SD. Statistics were performed using the ANOVA test: *p < 0.05, **p < 0.01, *** p < 0.001