Fig. 4

TREM2 expression and deglycosylation profiles in WT and p.R47H carriers. RIPA extracts from temporal cortices were used for the analyses. a. Western blot of a representative gel probed with C-terminal TREM2 antibody, Iba1, and actin antibodies. Three main TREM2 species are indicated: mature, immature and carboxy terminal fragment (CTF). b. The signal intensity in TREM2 species in WT and p.R47H samples (N = 16 for each group) was quantitated as previously described, and was plotted after normalization to actin and Iba1. The ratio between immature and mature TREM2 species and between CTF species normalized to TREM2 (full length: immature and mature species) were also plotted. c. Soluble fractions from RIPA buffer brain extracts were incubated in reaction buffers without enzymes (controls) or with endoH (1000U) or PNGaseF (1000U) overnight at 37 °C. The whole reaction mixture was analyzed by Western blot probed with a C-terminal TREM2 antibody. Blots containing two representative samples from WT and p.R47H brains are shown