Nanoliposomal-C6 ceramide inhibits the activation of ERK, which subsequently down-regulates survivin expression. (A) Western blot analysis was performed for p-ERK after treatment of RNK-16 cells with 5μM ghost or C6 nanoliposome for 18 hours. The equal loading of protein was confirmed by β-actin probing. (B) Western blot analysis was performed for p-ERK, total ERK, or survivin after treatment of NKL cells with different doses of PD098059 for 18 hours. The equal loading of protein was confirmed by β-actin probing. (C) Western blot analysis was performed for survivin after treatment of NKL cells with 25μM ghost (G) or C6 nanoliposome (C6) for 6, 16, and 24 hours. The equal loading of protein was confirmed by β-actin probing. (D) Western blot analysis was performed for survivin after treatment of RNK-16 cells with different doses of either ghost or C6 nanoliposome for 24 hours. The equal loading of protein was confirmed by β-actin probing. (E) Quantitative real-time RT-PCR was performed to measure levels of survivin mRNA after treatment of NKL cells with 25μM ghost or C6 nanoliposome for 6 and 16 hours. *P < .05 indicates significance between ghost and C6 nanoliposome treated samples (Student t test). (F) Quantitative real-time RT-PCR was performed to measure levels of survivin mRNA after treatment of RNK-16 cells with 5 and 20μM ghost or C6 nanoliposome for 24 hours. *P < .05 indicates significance between ghost and C6 nanoliposome-treated samples (Student t test).
