Trafficking of acetyl-C16-ceramide-NBD with long-term stability and no cytotoxicity into the Golgi
Tomohiko Makiyama, Hiroyuki Nakamura, Nobuo Nagasaka, Hisahiro Yamashita, Takuya Honda, Naoto Yamaguchi, Atsushi Nishida and Toshihiko Murayama
Accepted manuscript online: 23 JAN 2015 07:51AM EST | DOI: 10.1111/tra.12265
A549 cells were labeled with a new ceramide having an acetylated C-1 hydroxyl group, C16-N-acyl chain, and 4-nitrobenzo-2-oxa-1,3-diazole-bound C15-sphingosine (acetyl-C16-ceramide-NBD), and the washed cells were further cultured for 36 hr. The left and right panels show an overall picture and a typical picture of daughter cells, respectively. Acetyl-C16-ceramide-NBD preferentially accumulated in the Golgi complex before and after mitosis. Because of its weaker cytotoxicity and resistance to ceramide metabolic enzymes, acetyl-C16-ceramide-NBD is useful as a Golgi-labeling probe in various cell types.