Cellular apoptosis was quantified via TUNEL staining following the protocol of the One Step TUNEL Apoptosis Assay Kit (Keygen, Nanjing, China). The treated N2a cells and MCAO brain slices were stained according to the manufacturer’s instructions [36 (link)]. The survival rates of the neurons in the mouse penumbra were evaluated via Nissl staining. Briefly, the brain slices were stained with Nissl staining solution at 37°C for 10 min. Similarly, TUNEL-positive cells and typical Nissl bodies were counted under a fluorescence microscope (Olympus, Tokyo, Japan).
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