LPC-induced demyelination model of white matter injury

LPC-induced demyelinated lesions were generated in the ventral white matter of the spinal cord of 10-wk-old NG2-CreER-derived inducible conditional mutants. Tamoxifen (100 mg/kg of body weight) was administered five times daily by intraperitoneal injection at 4 wk prior to LPC-induced injury. We then performed the injection of 0.5 µL of 1% lysolecithin in the ventral white matter of the spinal cord via a pulled glass needle attached to the Hamilton syringe. Mice were harvested at multiple time points (between 3 d postlesion [dpl] and 10 dpl) and analyzed. Animals were perfused with 4% PFA. Spinal cords were dissected out and drop-fixed in 4% PFA overnight before being transferred into 20% sucrose. Tissue was embedded in OCT, sectioned, and stored at −80°C before in situ hybridization or immunofluorescence staining was performed as described above.