Fig 2: Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C).

Fig 3: MTX amplification and characterization of sDectin-1 cell pools.(A) Western blotting of supernatant samples from cultures adapted to different MTX concentrations using an mDectin-1 goat polyclonal antibody (1∶1000; AF1756; R&D Systems) with a HRP conjugated anti-goat antibody (1∶2000; Catalog number V8051; Promega). cHis pool and nHis pool are cell pools producing sDectin-1 with histidine tagged at the C- and N-terminals respectively. (B) Shake flask batch culture cell growth and sDectin-1 production profiles of sDectin-1 producing cell pools. Values shown represent mean values obtained from three replicate flasks. Error bars indicate the standard deviation of the experiment.

Fig 4: Enhanced expression of proteins associated with activated microglia in Grn −/−/Tmem106b −/− mice Western blot analysis of CD68, CLEC7A, TREM2, and ApoE in total brain lysates from 4.5‐month‐old mice with the given genotype. Asterisk indicates unspecific band (n = 3 biological replicates per genotype).Quantification of (A). Protein expression was normalized to levels in wild‐type animals. Data represent the mean ± SD.Expression of CD68 and the microglial marker IBA1 in sagittal brain sections. Representative images of indicated brain regions (TH, thalamus). Scale bar indicates 25 μm.Expression of CLEC7A and the microglial marker IBA1 in sagittal brain sections. Representative images of indicated brain regions (MY, medulla). Scale bar indicates 25 μm.Expression of TREM2 and the microglial marker IBA1 in sagittal brain sections. Representative images of indicated brain regions (MY, medulla). Scale bar indicates 25 μm.Data information: For statistical analysis of protein levels (B), one‐way ANOVA with Tukey's post hoc test was used to compare individual genotypes. Significance is indicated; *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001. Source data are available online for this figure.