Figure 2. sTREM2 promotes the production of inflammatory cytokines. (A) Primary microglia were treated with 4, 20, or 40 nM purified Fc or sTREM2-Fc fusion protein for 4 h. RNA was extracted, and the relative mRNA levels of IL-1β, IL-6, TNF, IL-10, Arg-1, and Ym-1 shown as bar graph were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 3 per group, one-way ANOVA). The mRNA level of 4 nM Fc-treated microglia served as a control. (B) Primary microglia isolated from WT or Trem2-KO were treated with 20 nM purified Fc or sTREM2-Fc fusion protein for 4 h. RNA was extracted and the relative mRNA levels of IL-1β, IL-6, and TNF shown as bar graphs were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 3 per group, unpaired Student’s t test). The mRNA level of Fc-treated WT microglia served as a control. (C) WT or Trem2-KO microglia were treated with 20 nM Fc or sTREM2-Fc fusion proteins for 24 h. Microglia were then stained by Alexa Fluor 488–labeled phalloidin. Representative photomicrographs are shown (blue, DAPI; green, phalloidin). Bar, 50 µm. (D) At least 55 cells from three independent experiments were selected to quantify the size of microglia, including area, diameter, and perimeter (unpaired Student’s t test). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Figure 2.

sTREM2 promotes the production of inflammatory cytokines. (A) Primary microglia were treated with 4, 20, or 40 nM purified Fc or sTREM2-Fc fusion protein for 4 h. RNA was extracted, and the relative mRNA levels of IL-1β, IL-6, TNF, IL-10, Arg-1, and Ym-1 shown as bar graph were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 3 per group, one-way ANOVA). The mRNA level of 4 nM Fc-treated microglia served as a control. (B) Primary microglia isolated from WT or Trem2-KO were treated with 20 nM purified Fc or sTREM2-Fc fusion protein for 4 h. RNA was extracted and the relative mRNA levels of IL-1β, IL-6, and TNF shown as bar graphs were determined by quantitative real-time PCR. β-Actin was used as an internal control (n = 3 per group, unpaired Student’s t test). The mRNA level of Fc-treated WT microglia served as a control. (C) WT or Trem2-KO microglia were treated with 20 nM Fc or sTREM2-Fc fusion proteins for 24 h. Microglia were then stained by Alexa Fluor 488–labeled phalloidin. Representative photomicrographs are shown (blue, DAPI; green, phalloidin). Bar, 50 µm. (D) At least 55 cells from three independent experiments were selected to quantify the size of microglia, including area, diameter, and perimeter (unpaired Student’s t test). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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