Journal: CNS Neuroscience & Therapeutics

Article Title: TREM2 Facilitates Myelin Debris Clearance but Exacerbates Chronic Inflammation and Fibrosis After Spinal Cord Injury

doi: 10.1002/cns.70777

Figure Lengend Snippet: TREM2 is primarily expressed on macrophages/microglia after SCI. (A) Representative immunostaining image of Iba1 (green), CD31 (green), PDGFRβ (green), or GFAP (green), TREM2 (red), and DAPI (blue) in sagittal sections at 7 dpi. Scale bars: Low magnification, 100 μm; high magnification, 20 μm. The asterisk indicates the lesion core. (B) Bar graphs show the percentage of each cell type expressing TREM2 from the mice described in (A). Asterisks show a statistically significant difference between macrophages/microglia and all other TREM2‐expressing cells. (C) Bar graphs show the percentage of TREM2‐expressing cells within each cell type from the mice described in (A). Asterisks show statistically significant differences in macrophages/microglia and all other TREM2‐expressing cells. (D) Representative immunostaining images of CX3CR1‐GFP (green), CCR2‐RFP (red), and TREM2 (white) in sagittal sections at 7 dpi. Scale bars: Low magnification, 100 μm; high magnification, 20 μm. The asterisk indicates the lesion core. (E) Bar graphs show the percentage of microglia and macrophages expressing TREM2 from the mice described in (D). Asterisks show a statistically significant difference between microglia and macrophages. (F) Bar graphs show the percentage of TREM2‐expressing cells within microglia and macrophages from the mice described in (D). Asterisks show statistically significant differences in microglia and macrophages. Statistical significance between experimental groups was calculated by one‐way ANOVA followed by Tukey's post hoc test (B, C), unpaired Student's t ‐test (E, F). *** p < 0.001, **** p < 0.0001. Data are presented as mean ± SEM; each point represents an individual mouse.

Article Snippet: The antibodies used in this study included the following: TREM2 (10 μg/mL, AF1729, R&D Systems, USA), PDGFRβ (5 μg/mL, AF1042‐SP, R&D Systems, USA), CD31 (1:100, AF3628, R&D Systems, USA), 5‐hydroxytryptamine (5‐HT) (1:5000, 20,079, Immunostar, Hudson, WI, USA), Iba1 (1:100, NB100‐1028, Novus, St Louis, MO, USA), CCR2 (1:100, NBP1‐48338, Novus, St Louis, MO, USA), fibronectin (1:100, 15613‐1‐AP, Proteintech, China), laminin (1:100, 23498‐1‐AP, Proteintech, China), glial fibrillary acidic protein (GFAP) (1:400, 13‐0300, Thermo Fisher Scientific, Waltham, MA, USA), CD68 (1:400, MCA1957, Bio‐Rad, Hercules, CA, USA), CSPG (1:200, C8035, Sigma‐Aldrich, USA), collagen I (1:200, ab21286, Abcam, USA), CST7 (1:100, PA5‐103772, Thermo Fisher Scientific, Waltham, MA, USA), IGF1 (1:100, ab223567, Abcam, USA), P2ry12 (1:500, EPR26298 ‐93, Abcam, USA), NeuN (1:100, EPR12763 , Abcam, USA), AKT (1:100, MA5‐41139, Thermo Fisher Scientific, Waltham, MA, USA), GSK3β (25 μg/mL, MAB2506, R&D Systems, USA), p‐AKT (1:100, sc‐514,032, Santa Cruz, USA), and p‐GSK3β (1:100, MA5‐14873, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Immunostaining, Expressing