Stereotactic Injection of ASO NEAT1 in Mice

Mice were anesthetized with ketamine 10% (100 mg/kg body weight) and xylazine 2% (10 mg/kg body weight) in 0.9% NaCl (max. injection volume 10 ml/kg body weight) and then fixed in a stereotactic frame on a warming plate under deep anesthesia. After application of an eye ointment, antiseptic treatment, skin incision, and drill hole trepanation (diameter: 0.6 mm), injections were made at x (1.5 mm from the sagittal suture), y (− 2.0 mm from the bregma), and z (2.5 mm depth) via a motorized injector (Stoelting Wood Dale, IL, USA). ASO NEAT1 (2 nmol, 2 μl) or ASO scramble (2 nmol, 2 μl) was slowly injected over 5 min using a 10 μl Hamilton cannula (outer diameter 240 μm; injection rate: 400 nl/min) once a week for 2 consecutive weeks before MCAO surgery (thus two injections in total). After the injection, the cannula remained in the tissue for 4 min to prevent reflux through the injection channel. The stereotactic injection paradigms essentially followed a protocol described by Jin et al. [17 (link)].