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Figure 2 | Molecular Cancer

Figure 2

From: Withanolide D induces apoptosis in leukemia by targeting the activation of neutral sphingomyelinase-ceramide cascade mediated by synergistic activation of c-Jun N-terminal kinase and p38 mitogen-activated protein kinase

Figure 2

WithaD induces ceramide production in K562 and MOLT-4 cells. (A) Cells were treated with WithaD for 0-2 hr and the intracellular ceramide content were determined using anti-ceramide antibody by flow cytometer as described in Materials and methods. Representative histogram (top) and the percentages of positive cells (bottom) are shown. Results are the mean ± S.D. in duplicate in three independent experiments. Asterisk indicates statistically significant difference (P < 0.005) in 2 hr treated cells with respect to untreated cells. (B) The intracellular ceramide levels induced by WithaD in K562 and MOLT-4 cells were measured in the cell lipid extract by the diacylglycerol kinase assay as described in Materials and methods. Cells were incubated with 1.5 and 0.5 μM WithaD respectively at the indicated times. * represents statistical significant difference in ceramide level between 15 min and control, where P < 0.05. ** indicates significant difference in ceramide level between 90 min and control. Data are expressed as % of control values and are the mean ± S.D. of three different experiments performed in triplicate. (C) Level of ceramide and SM were determined in K562 and MOLT-4 cells separated in HPTLC Si 60 plates after 1 hr of WithaD treatment, along with standard ceramide and SM. This is a representative profile of three independent experiments. (D) Densitometric quantitation of ceramide and SM in lipid extracts from treated and untreated K562 and MOLT-4 cells. Data are mean ± SD of three independent experiments.

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Molecular Cancer

ISSN: 1476-4598