Model 2004

The methods used to induce dissecting AAA have been described elsewhere 17 (link). Briefly, mice were treated with Ang II (1000 ng/kg per min; Sigma-Aldrich, MO, USA) or saline for 4 weeks via osmotic pumps (Model 2004; Durect Corporation, CA, USA) that were implanted subcutaneously in 16-week-old male WT and Plod1^-/- mice under anesthesia with 3% isoflurane in an O₂ mixture (1 L/min). The mice were observed daily for normal behavior or sudden death. The mice that died were immediately necropsied to determine the cause of death.
To investigate whether thrombospondin-1 is the key mediator of dissecting AAA progression in the absence of LH1, Plod1^-/-mice were intraperitoneally injected with 100 μL of TAX2 peptide (10 mg/kg) or scrambled control peptide (10 mg/kg) 3 times a week during the 4-week Ang II infusion period.
To further corroborate that LH1 is a potential critical therapeutic target for dissecting AAA, Plod1^-/-mice were intraperitoneally injected once with 2×10¹¹ AAV-LH1 or AAV-GFP particles in a total volume of 200 µl of supernatant. Three weeks later, the mice underwent Ang II infusion for 4 weeks.

Eight-week-old ApoE^–/– mice were implanted with a microosmotic pump (Model 2004, Durect Corporation, Cupertino, CA, USA), which was filled with 2.5 μg/kg/min Ang II (Sigma-Aldrich). Saline was used as a control for Ang II. The mice were given the treatment for 14 days. Forty-eight 8-week-old ApoE^–/– mice were randomly grouped as follows: saline infusion with RA (n = 6); Ang II infusion with RA (n = 17); Ang II infusion plus CIH (n = 16); and saline infusion plus CIH (n = 9). Two weeks after CIH, mice were infused with Ang II for another 2 weeks in the CIH groups.