Skip to main content
Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: Functional involvement of γ-secretase in signaling of the triggering receptor expressed on myeloid cells-2 (TREM2)

Fig. 1

Inhibition of γ-secretase impairs TREM2-dependent Ca2+ signaling. a COS7 cells were transfected with mouse myc-TREM2-FL-GFP or control vector encoding GFP only. Constructs encoding TREM2 and DAP12 have been described earlier [18]. TREM2 was always co-transfected with DAP12 to allow efficient subcellular transport [15, 22]. Expression of TREM2-FL and TREM2-CTF was analyzed by Western immunoblotting using either anti-myc or anti-GFP antibodies. The γ-secretase inhibitor DAPT (1 μM, 18 h pretreatment) significantly increased the ratio of TREM2-CTF to TREM2-FL (***p < 0.001; n = 3). b Ratiometric determination of intracellular Ca2+ levels upon anti-myc antibody induced TREM2 activation in the absence or presence of DAPT (1 μM, 18 h pretreatment). COS7 cell treatment with DAPT attenuates the TREM2-evoked increase in intracellular [Ca2+] (****p < 0.0001; n = 4–6). c Morphological change upon activation of TREM2 in the absence or presence of DAPT. COS7 cells co-expressing mouse myc-TREM2-GFP and DAP12 were seeded onto coverslips and monitored by light microscopy for 10 min to determine spontaneous changes of the cell area. After 10 min, monoclonal anti-myc antibodies were added to the medium and cells were imaged for additional 2 h. Changes in cell surface area were determined by subtraction of the area after 2 h of stimulation from that before stimulation. Activation of TREM2 with anti-myc antibody induced a significant decrease in cell size (*p < 0.05). The inhibition of γ-secretase with DAPT prevented the TREM2-induced shrinkage of cells (*p < 0.05; n = 3). d COS7 cells were transfected with mouse myc-TREM2-FL-GFP and treated with anti-myc antibody for 2 h after overnight incubation with or without DAPT. TREM2 FL and CTFs were detected by western immunobloting

Back to article page