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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Nat Cell Biol. 2015 Jan 12;17(2):170–182. doi: 10.1038/ncb3090

Figure 3. TAMs in human primary GBMs and xenografts are monocyte-derived macrophages from peripheral blood.

Figure 3

a, Immunofluorescent staining of CCR2 (the marker for monocyte-derived macrophages) and CX3CR1 (the microglia marker) in GBM xenograft and normal brain tissue. Frozen sections of GSC-derived tumor (T387) and the adjacent normal mouse brain were immunostained with antibodies against CCR2 (green) and CX3CR1 (red) and counterstained with DAPI (blue). CCR2+ cells were detected only in tumor tissue, while CX3CR1+ cells were detected only in normal brain. Scale bar, 40μm. b, Immunohistochemical staining of CCR2 and CX3CR1 in human primary GBMs and normal brain tissue. Two consecutive tissue microarray slides (US Biomax) containing GBMs and normal brain tissues were immunostained with antibody against CCR2 or CX3CR1 (brown) and counterstained with hematoxylin. GBM tumors showed abundant CCR2+ cells (the monocyte-derived TAMs) but not CX3CR1+ cells (microglia), while normal brain tissues contained CX3CR1+ cells but not CCR2+ cells. Scale bar, 40μm. c, Graphical analysis of (b) in tissue microarrays showed that CCR2+ cells (monocyte-derived TAMs) were detected in the majority (82.6%) of GBM cases (data from 23 tumors) and the minority (20%) of normal brains (data from 5 normal samples). In contrast, CX3CR1+ cells (microglia) were detected in all normal brains but only 4.3% of GBM tumor cases. d, Immunofluorescent staining of Iba1 and the CD31 in human primary GBMs. Frozen sections of a primary GBM (CW2445) were immunostained with antibodies against Iba1 to label TAMs (green) and CD31 to mark vessels (red) and counterstained with DAPI (blue). Abundant TAMs are enriched in perivascular niches. Scale bar, 80μm. e, Immunofluorescent staining of the TAM marker Iba1 and the endothelial marker Glut1 in GSC-derived xenografts. Frozen sections of GBM xenografts derived from GSCs (T387) were immunostained with antibodies against Iba1 (green) and Glut1 (red) and counterstained with DAPI (blue). TAMs (green) were localized near vessels but not in the area lacking blood vessel. Scale bar, 80μm.