Fig. 1

Schematic of experimental flow for rats and mice cortical slices. Coronal slices of mouse and rat occipital cortex are pre-incubated ex vivo with either a vehicle or AKU-005 (100 nM). For live calcium imaging, one slice per treatment is used and then exposed to either a vehicle or high KCl (100 mM) to induce cortical spreading depression (CSD). The remaining slices undergo the same ex vivo treatments of high KCl or vehicle without imaging. All slices from the same animal and treatment group are then frozen together, homogenized, and divided into two portions. One portion is utilized for activity-based protein profiling (ABPP) assay, during which additional in vitro inhibitory treatments are performed. The remaining homogenized material is analyzed using mass spectrometry