Three human recombinant ApoE proteins, isoforms 2, 3 and 4 (PeproTech, Rocky Hill, NJ, USA), were separately added into Neurobasal/B27 to culture the explants of ApoE-/- mice after planting. The ApoE final concentration was 10 µg/ml in medium, which is similar to the concentration of ApoE in human cerebrospinal fluid (CSF)22 (link)–24 (link, link) . In addition, after axonal transection, the explants of the ApoE-/- mice were continuously cultured in Neurobasal/B27 with human recombinant ApoE2/3/4 proteins for 24 h.
JNK/ERK/p38 inhibitors, which included SP600125, U0126, and SB203580 (Beyotime, Shanghai, China), were also used as interventional means. The final concentration of the three inhibitors was 10 μM, and they were dissolved in dimethyl sulfoxide (DMSO, Beyotime)25 (link),26 (link). In the control group, only DMSO was added without the inhibitors. After explant planting, the explants were separately treated by medium with the JNK, ERK and p38 inhibitors for 24 h. Subsequently, the media were replaced with fresh media without the inhibitors27 (link).
JNK/ERK/p38 inhibitors, which included SP600125, U0126, and SB203580 (Beyotime, Shanghai, China), were also used as interventional means. The final concentration of the three inhibitors was 10 μM, and they were dissolved in dimethyl sulfoxide (DMSO, Beyotime)25 (link),26 (link). In the control group, only DMSO was added without the inhibitors. After explant planting, the explants were separately treated by medium with the JNK, ERK and p38 inhibitors for 24 h. Subsequently, the media were replaced with fresh media without the inhibitors27 (link).
