SEARCH

SEARCH BY CITATION

FilenameFormatSizeDescription
nan12086-sup-0001-si.tif16181K

Figure S1. Effect of human IL-34 on the differentiation of human ML cells. (A) Fluorescent microscopic images of human monocytes cocultured with human astrocytes in the presence or absence of M-CSF (50 ng/ml) or IL-34 (50 ng/ml). Data are representative of three independent experiments. Bar: 100 μm. (B) Dose–response relationship between M-CSF/IL-34 and the number of ML cells differentiated from monocytes (cell number per mm2). Data are presented as mean ± SD. *P < 0.05 as compared with untreated cells. (C) Immunofluorescence of human ML cells. Human monocytes culture with human IL-34 (50 ng/ml) mainly showed spindle-like morphology (arrow) and all cells were positive for TREM2. Data are representative of three independent experiments. Bar: 50 μm.

nan12086-sup-0002-si.tif6804K

Figure S2. Functional characterization of isolated ML cells. (A) Morphology of peritoneal macrophages, microglia isolated from mixed glial culture, and murine ML cells isolated from astrocyte coculture. Data are representative of three independent experiments. Bar: 50 μm. (B) Immunofluorescence of isolated murine ML cells. All cells were positively stained with Iba1, CD11b, TREM2 and MHC II. Data are representative of three independent experiments. Bar: 100 μm. (C) Phagocytosis assay: percentage of peritoneal macrophages, microglia and murine ML cells phagocytosing zymosan-coated particles. Data are presented as mean ± SD. (D) Cytokine production after LPS treatment. Cells were treated with or without LPS (20 ng/ml) and concentration of TNFα and IL-6 in culture supernatants were determined by ELISA. Incubation periods were 6 h for TNFα and 24 h for IL-6. Data are presented as mean ± standard error (SE).

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.