Figure 4. The AD risk-associated variants impair the functions of sTREM2 in microglia. (A) The purified Fc, sTREM2 WT-Fc (WT), sTREM2 R47H-Fc (R47H), and sTREM2 R62H-Fc (R62H) fusion proteins were analyzed by silver staining. (B) Microglia obtained from WT or Trem2-KO mice were cultured with 20 nM Fc, WT, R47H, or R62H protein for 24 h after GM-CSF withdrawal. TUNEL staining was then performed and 12 different fields from three independent experiments were selected to quantify the number of TUNEL-positive cells (one-way ANOVA). (C) Primary microglia from WT mice were treated with 20 nM Fc, WT, R47H, or R62H protein for 4 h. RNA was extracted and the relative mRNA levels of IL-1β, IL-6, and TNF shown as bar graph were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 5 per group, one-way ANOVA). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Figure 4.

The AD risk-associated variants impair the functions of sTREM2 in microglia. (A) The purified Fc, sTREM2 WT-Fc (WT), sTREM2 R47H-Fc (R47H), and sTREM2 R62H-Fc (R62H) fusion proteins were analyzed by silver staining. (B) Microglia obtained from WT or Trem2-KO mice were cultured with 20 nM Fc, WT, R47H, or R62H protein for 24 h after GM-CSF withdrawal. TUNEL staining was then performed and 12 different fields from three independent experiments were selected to quantify the number of TUNEL-positive cells (one-way ANOVA). (C) Primary microglia from WT mice were treated with 20 nM Fc, WT, R47H, or R62H protein for 4 h. RNA was extracted and the relative mRNA levels of IL-1β, IL-6, and TNF shown as bar graph were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 5 per group, one-way ANOVA). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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