Antagonistic TREM2 antibodies reduce hyperactivation of PGRN deficient hiMGLA.Figure 5A. Expression of all analyzed gene transcripts in GRN-/- hiMGL treated with control isotype antibody, Ab1 or Ab2 in comparison to WT hiMGL. Data show the mean of four individual treatments and NanoString measurements. The mRNA counts for each gene were normalized to the mean value of all WT samples followed by a log2 transformation. B. Volcano plot presentation of the differently expressed transcripts in GRN-/- hiMGL treated with isotype compared to WT hiMGL. Genes with more than 20% significantly changed expression are marked in purple (upregulated) or blue (downregulated). Data information: Data represent mean ± SEM. For statistical analysis the unpaired, two-tailed student`s t-test was performed, Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant. C. Volcano plot presentation of the differently expressed transcripts in GRN-/- hiMGL treated with Ab1 comparison to WT hiMGL. Genes with more than 20% significantly changed expression are marked in purple (upregulated) or blue (downregulated). Data information: Data represent mean ± SEM. For statistical analysis the unpaired, two-tailed student`s t-test was performed, Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant. D. Volcano plot presentation of the differently expressed transcripts in GRN-/- hiMGL treated with Ab2 comparison to WT hiMGL. Genes with more than 20% significantly changed expression are marked in purple (upregulated) or blue (downregulated). Data information: Data represent mean ± SEM. For statistical analysis the unpaired, two-tailed student`s t-test was performed, Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant. E. Transcript levels of DAM gene transcripts significantly altered in GRN-/- hiMGL, treated with Ab1 or Ab2 in comparison to isotype treatment from the data set in A, normalized to the mean of the WT hiMGL samples (n=4, biological replicates). Data information: Data represent mean ± SEM. For statistical analysis one-way ANOVA with Dunnett`s post hoc test was used to compare Ab1 and Ab2 (20 µg/ml and 40µg/ml) conditions to the isotype treated condition, Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant. F. Transcript levels of CTSD, NPC2 and CD68 of WT and GRN-/- hiMGL untreated, treated with isotype control, and Ab1 or Ab2 in comparison to WT hiMGL from the data set in A normalized to the mean of the WT hiMGL samples (n=4, biological replicates). Data information: Data represent mean ± SEM. For statistical analysis one-way ANOVA with Dunnett`s post hoc test was used to compare Ab1, Ab2 (20 µg/ml and 40µg/ml) and isotype treated condition to WT cells. Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant. G. Catalytic activity of cathepsin D (CatD) in untreated WT and GRN-/- hiMGL or GRN-/- hiMGL treated with isotype control, Ab1 or Ab2 (20 μg/ml, 40 μg/ml), as measured by a CatD in vitro activity assay (n=3, biological replicates). Data information: Data represent mean ± SEM. For statistical analysis one-way ANOVA with Dunnett`s post hoc test was used to compare Ab1, Ab2 (20 µg/ml and 40µg/ml) and isotype treated condition to WT cells. Statistical significance was set at *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, *****, p < 0.00001, and ns, not significant.