Lipid analyses

Plasma was collected from five-hour fasted mice preceding AAV injection and at 4, 8, 12, 16, and 20 weeks post-injection. Total plasma cholesterol at all time points was measured using the Wako Cholesterol E kit (999-02601). Gel filtration chromatography was completed with 220 μL pooled plasma from mice with high plasma cholesterol. Pooled plasma was loaded into a 200 μL loop on an Amersham-Pharmacia ÄKTA chromatography system equipped with two Superose HR6 columns in tandem and eluted with TBS at a flow rate of 0.5 mL/min²². Lipoprotein cholesterol and triglyceride levels were determined using the Wako Cholesterol E (999-02601) and Infinity Triglycerides (TR22421) kits using 100 μL of each 1 mL fraction and expressed as micrograms per milliliter.