Extended Data Fig. 2: Generation and validation of cell specific cholesterol synthesis mutants.

a, Quantitative RT-PCR analysis determining the expression of housekeeping reference genes in corpus callosum or isolated cells from mice (n = 4 animals) treated with cuprizone (A astrocytes, EC endothelial cells, M microglia/macrophages, OL oligodendrocytes). b, Conditional mutants were generated by crossing SQSflox/flox mice to Cre driver lines. Fdft1 was inactivated by tamoxifen injections, and mice were analyzed at time points as indicated (middle panel). Fdft1 (SQS) expression in acutely isolated cells targeted by the conditional mutation shown as mean of fold values (OLcKO n = 4, OPCcKO n = 3, AcKO n = 3, McKO n = 3, ECcKO n = 3) normalized to cells from control mice (OLcKO-Ctrl n = 4, OPCcKO-Ctrl n = 3, AcKO-Ctrl n = 3, McKO-Ctrl n = 4, ECcKO-Ctrl n = 3; set to 1). Asterisks mark significant changes (Student’s t-test, two-sided). c, Body weight (OLcKO n = 15, OPCcKO n = 7, AcKO n = 5, McKO n = 10, ECcKO n = 5; OLcKO-Ctrl n = 11, OPCcKO-Ctrl n = 6, AcKO-Ctrl n = 5, McKO-Ctrl n = 9, ECcKO-Ctrl n = 6) and total serum cholesterol (OLcKO n = 7, OPCcKO n = 7, AcKO n = 5, McKO n = 5, ECcKO n = 8; OLcKO-Ctrl n = 4, OPCcKO-Ctrl n = 6, AcKO-Ctrl n = 5, McKO-Ctrl n = 5, ECcKO-Ctrl n = 5) of analyzed cell-specific cholesterol synthesis mutants in (b) at the age of 22 weeks. Bars represent mean of individual animals. Asterisks mark significant changes (Student’s t-test, two-sided). d, Evaluation of astrocyte-specific cholesterol synthesis mutants (Aldh1l1-CreERT2 driver line, Winchenbach et al., 2016), determining Fdft1 expression in isolated astrocytes, body weight and total serum cholesterol in mutants (isolated astrocytes n = 3 animals; body weight n = 9 animals, cholesterol n = 7 animals and Cre controls (isolated astrocytes n = 4 animals; body weight n = 3 animals, cholesterol n = 8 animals). Bars represent the means with individual data points normalized to controls. Asterisks mark significant changes (Student’s t-test, two-sided). e, Targeted sterol metabolism gene expression profile of isolated cells from the corpus callosum of n = 4 untreated WT animals. The relative expression heat map shows low transcript levels (blue) and high transcript levels (red) for each gene normalized to the mean of all samples. f, Quantification of histological stainings for myelination (Gallyas), mature oligodendrocytes (CAII), oligodendrocyte linage cells (Olig2), microgliosis (MAC3) and astrogliosis (GFAP) in Aldh1l1-CreERT2 astrocyte cholesterol synthesis mutants (n = 4 animals) as in (d) at 6 weeks cuprizone. Bars represent the means with individual data points normalized to controls (n = 4 animals) set to 100). ***p < 0.001, **p < 0.01, *p < 0.05.