Figure S2.

Defective cholesterol esterification affects remyelination, inflammation, and cholesterol crystal formation. (A) TEM of 21-dpi lesions of WT and ACAT KO mice showing lipid droplets and remyelinated axons (yellow arrows) in WT but not in KO cells. (B) The g-ratio calculations of 21-dpi lesions of WT and ACAT KO mice. (C and D) Percentage of IBA1⁺ cells, which are also MAC2⁺, and quantification of the density of astrocytes (labeled for GFAP) in 4- and 21-dpi lesions of WT and ACAT KO mice. (E) Quantification of crystal intensity in 21-dpi lesions of WT and ACAT KO mice by confocal reflection microscopy. (F–H) Confocal images of WT microglia and quantification of crystal colocalization with lipid droplets labeled with PLIN2 (yellow) and LAMP1⁺ lysosomes (red). Examples of colocalization with lysosomes and lipid droplets are highlighted by arrowheads and arrows, respectively. (I) Representative TEM pictures of cholesterol crystals in WT and ACAT KO lesions at 21 dpi. WT microglia occasionally showed small crystals (<5 μm, magenta arrowheads), while large crystals (>15 μm) were rarely observed in ACAT KO lesions (magenta arrows). The results are representative of three independent experiments. Scale bars, 2 μm in A, 3 μm in C, and 5 μm in I. *, P < 0.05; **, P < 0.01; ***, P < 0.001. CC, cholesterol crystals; IDA, integrated density area.