Figure 1. sTREM2 promotes cellular viability and suppresses cellular apoptosis in microglia. (A) Schematic representation of full-length human TREM2 and sTREM2-Fc fusion proteins. SP, signal peptide; ECD, extracellular domain; TM, transmembrane domain. (B) The purified Fc and sTREM2-Fc fusion proteins were analyzed by silver staining. (C) The cell viability of WT and Trem2-KO microglia were analyzed after cultured with Fc (20 nM), sTREM2-Fc fusion protein (20 nM), or GM-CSF (25 ng/ml) for 24 h (n = 4 per group, one-way ANOVA). The cell viability of Fc-treated WT microglia served as a control. (D) The cell viability of WT and Trem2-KO microglia were analyzed after culture with 20 nM active (native) or inactive (heat-inactivated for 1.5 h) Fc or sTREM2-Fc protein for 24 h after GM-CSF withdrawal (at least three repeats per group, unpaired Student’s t test). The cell viability of active Fc-treated WT microglia served as a control. (E) The proliferation of WT and Trem2-KO microglia were analyzed after cultured with Fc (20 nM), sTREM2-Fc protein (20 nM) or GM-CSF (25 ng/ml) for 24 h (n = 4 per group, one-way ANOVA). The BrdU incorporation rate of Fc-treated WT microglia served as a control. (F) Microglia from WT or Trem2-KO mice were cultured with 20 nM active (native) or inactive (heat-inactivated for 1.5 h) Fc or sTREM2-Fc 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 (unpaired Student’s t test). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.
Figure 1.

sTREM2 promotes cellular viability and suppresses cellular apoptosis in microglia. (A) Schematic representation of full-length human TREM2 and sTREM2-Fc fusion proteins. SP, signal peptide; ECD, extracellular domain; TM, transmembrane domain. (B) The purified Fc and sTREM2-Fc fusion proteins were analyzed by silver staining. (C) The cell viability of WT and Trem2-KO microglia were analyzed after cultured with Fc (20 nM), sTREM2-Fc fusion protein (20 nM), or GM-CSF (25 ng/ml) for 24 h (n = 4 per group, one-way ANOVA). The cell viability of Fc-treated WT microglia served as a control. (D) The cell viability of WT and Trem2-KO microglia were analyzed after culture with 20 nM active (native) or inactive (heat-inactivated for 1.5 h) Fc or sTREM2-Fc protein for 24 h after GM-CSF withdrawal (at least three repeats per group, unpaired Student’s t test). The cell viability of active Fc-treated WT microglia served as a control. (E) The proliferation of WT and Trem2-KO microglia were analyzed after cultured with Fc (20 nM), sTREM2-Fc protein (20 nM) or GM-CSF (25 ng/ml) for 24 h (n = 4 per group, one-way ANOVA). The BrdU incorporation rate of Fc-treated WT microglia served as a control. (F) Microglia from WT or Trem2-KO mice were cultured with 20 nM active (native) or inactive (heat-inactivated for 1.5 h) Fc or sTREM2-Fc 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 (unpaired Student’s t test). Data represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

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