Journal: Cell chemical biology

Article Title: TREM2 is thyroid hormone regulated making the TREM2 pathway druggable with ligands for thyroid hormone receptor.

doi: 10.1016/j.chembiol.2021.07.014

Figure Lengend Snippet: Figure 4. T3 and sobetirome stimulate and NH-3 blocks phagocytosis by microglia (A–D’) C57BL/6 mouse primary microglia cells in culture were treated with DMSO vehicle (A + A’), 10 nM T3 (B + B’), 1 mM sobetirome (C + C’), or 2 mM NH-3 (D + D’) for 24 h before 2 mm diameter fluorescent beads (100/cell) were introduced 2 h before cells were fixed and stained for CD11B (green). (E) Elongated morphology of an unactivated/ramified microglia cell. (F) Morphology of an activated microglia cell upon treatment with sobetirome (see the supplemental information). (G) Three-dimensional view down three separate axes for visualization of beads inside the uppermost cell in the sobetirome treatment group (image generated using Imaris Section function). (H) Quantification of phagocytosed beads per treatment group. Scale bars, 20 mm (A–D), 10–20 mm (A’–D’), and 8–10 mm (E–G), as noted. Statistical significance was determined by a two-tailed, unpaired t test for comparisons between vehicle and group then were plotted together. Asterisks represent significant difference from vehicle unless otherwise noted: *p % 0.05, **p % 0.01, ***p % 0.001. All graphs show mean ± SEM.

Article Snippet: The cells were then permeabilized with PBS containing 0.05% saponin (saponin was included in all subsequent incubations and washes) and stained for microglial marker CD11b using monoclonal rat anti-mouse CD11b (1:200 dilution, AbD Serotec, #MCA711) in conjunction with Alexa Fluor 488-conjugated donkey anti-mouse secondary antibody (1:400 dilution,Thermo Fisher, A21202).

Techniques: Staining, Generated, Two Tailed Test