Conditional Glut1 Knockout in Mice

Study subjects were maintained on a 129 S6/SvEvTac genetic background. Animals bearing the Glut1^Δ allele were generated as previously described (16 (link)). Mice harboring the Tie2-Cre (stock no. 008863), Zp3-Cre (stock no. 003651), and CreER (stock no. 008463) drivers were obtained from the Jackson Laboratory, backcrossed over 6 generations to the 129 S6/SvEvTac genetic strain background and genotyped using primers described in Supplementary Information (SI). Note that although the Tie2-Cre line is an established tool to deplete proteins in ECs, it is also reported to express Cre in a limited subset of non-ECs, notably macrophages. TM administrations first involved preparing stock solutions (20 mg/mL or 50 mg/mL) of the compound (T5648, MilliporeSigma) in 100 μL of ethanol and 900 μL corn oil. Three doses of 250 mg/kg (to 2-week-old mice) and 450 mg/kg (to 8-week-old mice) were delivered by oral gavage over consecutive days to the mice. P2 mice received 2 doses (125 mg/kg) of the TM on consecutive days. Subsequent analyses were then conducted at either 2 weeks of age (brain microvasculature studies) or at approximately 5 months of age (remaining assessments).