Figure 4 | Scientific Reports

Figure 4

From: E2F signature is predictive for the pancreatic adenocarcinoma clinical outcome and sensitivity to E2F inhibitors, but not for the response to cytotoxic-based treatments

Figure 4

Characterization of the ly101-4B, a compound with an anti-E2F activity. (a) Molecular structure of the ly101-4B a triazole nucleoside compound. (b) Luciferase activity assay using the 6x E2F-luciferase reporter in MiaPaCa2 cell line treated during 72 h with ly101-4b (25 µM) or vehicle treated in control. (c) MiaPaca2 cell lines were treated as in b. Cell viability was assessed using the PrestoBlue reagent for 3 h incubation at 37 °C according to the manufacturer. Fluorescences values were blank substracted and normalized to control (vehicle treated) (d) Effects of ly101-4B on caspases 3 and 7 proteolytic activities. MiaPaca2 cells were treated 72 h with ly101-4b (25 µM). Results are expressed relative to control values (=1). Data are mean ± SEM of tree independent experiments. (e) Effect of ly101-4B on cytosolic LDH release. MiaPaca2 cells were treated 48 h with ly101-4B. Results are expressed as percentage relative to vehicle treated. Data are mean ± SEM of three independent experiments of different cultures, each one performed in triplicates. (f,g) MiaPaCa2 cells in exponential growth phase were treated with ly101-4B for 12 h and then labeled with [3H] Thymidine (10 μCi/ml) in f or [3H] Uridine (10 μCi/ml) in g for a 6 h period. In f, treatment with actinomycin d was used as a negative control of RNA synthesis. DNA and RNA synthesis rates were determined according to the radioactivity by using liquid scintillation counting. Each experiment was performed in triplicate. Data are expressed in percentage relative to vehicle treated condition.

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