Figure 2 | Cell Death & Disease

Figure 2

From: C6-ceramide synergistically potentiates the anti-tumor effects of histone deacetylase inhibitors via AKT dephosphorylation and α-tubulin hyperacetylation both in vitro and in vivo

Figure 2

In vitro synergism between TSA and C6-ceramide in inducing cancer cell apoptosis. The apoptosis of CaOV3 cells with indicated treatment (control, 100 ng/ml of TSA, 10 μg/ml of C6-ceramdie and TSA plus C6-ceramide) for 32 h was detected by different methods including TUNEL assay (a), Hoechst 33342 assay (b), Histone DNA apoptosis ELISA assay (d). For each Hoechst experiment, at least 200 cells in 10 random scope fields were counted for apoptotic rate (c). CaOV3 cells were treated with 100 ng/ml of TSA in the presence or absence of 10 μg/ml of C6-ceramide for 8 and 16 h, the expression level of Bax, cleaved-caspase 3, -9 and β-actin were detected by western blot (e). CaOV3 cells were pretreated with caspase inhibitors Z-DEVDfmk (50 μM) or z-LEHDfmk (50 μM) for 2 h, followed by TSA (100 ng/ml) plus C6-ceramide (10 μg/ml) treatment; cell death was detected by MTT assay (f) at 48 h after treatment, and CaOV3 cells apoptosis was detected by TUNEL assay 36 h after treatment (g). CaOV3 (h) were exposed to HDACIs sodium butyrate (SB, 1 mM) or suberoylanilide hydroxamic acid (SAHA, 1.5 μM), in the presence or absence of C6-ceramide (10 μg/ml) for 36 h, cell apoptosis was detected by Histone DNA apoptosis ELISA assay. Data are presented as the mean±S.D. for three independent experiments. Experiments in this figure were repeated at least three times and similar results were obtained. Bar=10 μm. *P<0.05 versus that indicated in the figure (ANOVA)

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