Zp3 cre

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).

The floxed Nat10 (Nat10^lox/lox) alleles were generated by GemPharmatech Co., Ltd. Conditional Nat10 knockout mice were achieved by crossing Nat10^lox/lox mice with Stra8-GFPCre and with Zp3-Cre mice to attain the Nat10-ScKO and Nat10-ZcKO offspring, respectively. Nat10 KO MEF cells were achieved from the embryo of Nat10-Ubc mice by crossing Nat10^lox/lox mice with Ubc-CreERT2 KI mice. The Stra8-GFPCre knock-in (KI) mouse line was generated in Ming-Han Tong’s Lab at the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences. Zp3-Cre and Ubc-CreERT2 KI mice were obtained from Jackson Laboratory. All mice were from the C57BL/6J background and were bred in a specific pathogen-free (SPF) facility with a 12 h light/dark cycle and with free access to food and water. All animal experiments were approved by the Animal Care and Research Committee of the University of Science and Technology of China (USTC).