C16-Ceramide 

C16-Ceramide 是一种天然小分子，通过直接选择性结合激活 p53。

C16-Ceramide Preparation, Cell Incubation, and Animal Injection.

C16-ceramide was dissolved in dimethyl sulfoxide (DMSO) and conjugated with 1−2% (w/v) fatty acid-free bovine serum albumin (BSA) by vortexing. Hereafter it is called the C16-ceramide mixture. Primary cortical neurons were washed with serum-free DMEM once, which was followed by treatment with different concentrations or timings of the C16-ceramide mixture. We incubated the cells forming control group with serum-free DMEM containing vehicle solvent plus 1% to 2% fatty acid-free BSA. The vehicle DMSO content was kept to <0.1% (v/v) of the total amount so as not to interfere with effect of reagents.

In order to avoid any possible undesirable effect of DMSO on the mice, C16-ceramide dissolved in mineral oil was delivered into mice via subcutaneous injection 5 days a week for 4 weeks. The dose was adopted 0.18 μg/g body weight on the basis of the plasma concentration reported by Ichi et al. Actually, double-dose 0.36 μg/g body weight was also adopted. The mice were weaned at 3 weeks old and then injected with either C16-ceramide dissolved in mineral oil or mineral oil until they were 7 weeks old.