Fig 1: Glioblastoma stem cells (GSCs) induce macrophage toward M2 polarization which infiltrates more in GBM. (A) Representative immunofluorescent (IF) staining of M2-like TAMs marker (CD163 and CD206) in human GBM tissues. Nuclei were counterstained with DAPI (blue). Scale bar represents 20 µm. (B) Kaplan–Meier survival plots of M2-like TAMs signature score showed a higher score indicated a poorer prognosis. p ? 0.001, log-rank test. (C) The expression levels of M0 markers (CD11b and CD68) were determined by RT-qPCR. (D, E) Conditioned media of GSC promoted macrophage migration through transwell filters. (F) After PMA induction, THP1 (Mf) were cocultured with GSCs. The expression levels of M2 markers (CD163, Arg1, and IL10) and M1 markers (CD80 and iNOS) were determined by RT-qPCR. (G) THP1 (Mf) were cocultured with GSCs (GSC11 or GSC23). Flow cytometry was applied to measure macrophage marker (CD11b) and M2 macrophage-related phenotypic marker (CD163) on the surfaces of macrophages. (H) The ratio of CD11b-positive plus CD163-positive (CD11b+CD163+) macrophages were quantitated. (I, J) Macrophages were cocultured with GSCs treated with DMSO or exosome secretion inhibitor GW4869. Flow cytometry and quantification were performed to analyze the proportion of CD11b+CD163+ macrophages. Data depict the mean ± standard deviation and are representative of three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).
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