Fig. 8

EED inhibitors hinder the chemotaxis of human moDCs. A Scheme of human moDCs culture and inflammation-associated phenotype detection. B The morphological changes from monocytes (Day0) to immature moDCs (Day 6) and eventually LPS-challenged mature moDCs (Day7). Scale bar: 20 μm. n = 3. C Representative images (left) and bar graph (right) for flow cytometry analysis of the frequency of CD209⁺ moDCs after cell culture for 7 days. n = 3. D Western blotting analysis (left) and its corresponding quantification (right) for the expression of EED and H3K27me3 in human moDCs treated with A395 (5 μM), MAK683 (5 μM), EEDi-5285 (5 μM), EED226 (10 μM) and LPS (300 ng/mL) for 24 h. n = 3. E qRT-PCR analyses of the mRNA levels of WNT-related genes in human moDCs treated with EEDi-5285 (5 μM), EED226 (10 μM) and LPS (300 ng/mL) for 24 h. n = 3. F, G Representative images (E) and statistics (F) for transwell analysis in human moDCs after treating with A395 (5 μM), MAK683 (5 μM), EEDi-5285 (5 μM), EED226 (10 μM) and LPS (300 ng/mL) for 24 h. Scale bar: 200 μm. n = 3. H, I Representative images (H) and bar graph (I) for flow cytometry analysis of the expression of CD83 and HLA-DR (MHCII) in CD209⁺ moDCs after treatment with A395 (5 μM), MAK683 (5 μM), EEDi-5285 (5 μM), EED226 (10 μM) and LPS (300 ng/mL) for 24 h. n = 3. J Elisa of TNF-α and IL-10 levels in the supernatant of human moDCs after treatment with A395 (5 μM), MAK683 (5 μM), EEDi-5285 (5 μM), EED226 (10 μM) and LPS (300 ng/mL) for 24 h. n = 3. Data are presented as mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001