Fig. 6

Trem2 H157Y increases TREM2 shedding in 5xFAD mice. A. TREM2 in TBS and TBSX lysates from our mouse models and Trem2-KO mouse were detected using an N-terminal antibody (5F4) through Western blotting. B-C. TREM2 levels in TBS (B) and TBSX (C) were quantified and normalized to WT for each genotype. Bands indicated below 37kD were quantified. D. Full-length TREM2 levels were measured in the brain tissues from mice of each genotype. E. Soluble TREM2 levels were measured in the brain tissues from mice of each genotype. F. Ratios of soluble versus full-length (s/fl) TREM2 amount were calculated and normalized to WT mice. G. Ratios of s/fl TREM2 amount were calculated for TREM2-WT and TREM2-H157Y and normalized to TREM2-WT in Het mice. D-G, N = 3 mice/sex/ genotype at 8.5 months of age, mixed sex. Data are presented as Mean ± SEM. One-way ANOVA with multiple comparisons was used in D-F with Welch's correction. Wilcoxon matched-pairs signed rank test was applied to G. H. Serum TREM2 was examined by ELISA for each genotype. I. SYK, pSYK and actin were detected in the RIPA lysates of isolated microglia from WT and Hom mice. J-K. pSYK (J) and SYK (K) were quantified and normalized to WT. L. Ratios of pSYK/SYK were calculated and normalized to the WT mice. A-C, H. N = 19–24 mice per genotype at 8.5 months of age, mixed sex. Kruskal–Wallis tests with uncorrected Dun’s multiple comparisons were used. I-L.N = 3 mice/sex/genotype at 8.5 months of age. Unpaired t-tests were used. Data are presented as Mean ± SEM. N.S., not significant. * p < 0.05. **p < 0.01. ***p < 0.001