Fig. 6. Evaluation of inhibitory activity of oligo-NSAs in cellulo. (a) Intracellular protein levels of p53, p21, and β-actin in MDM2-overexpressing SJSA-1 cells treated with oligo-NSAs. SJSA-1 cells were incubated with 0, 2.5, 5, or 10 μM 5, 10 μM 1, 10 μM 5-Rv, or 10 μM Nutlin-3a for 8 h and cell lysates were analyzed by western blotting. Representative results of three independent experiments are shown. Other results are shown in Fig. S28.† (b) Measurements of apoptotic cells by staining with Annexin V-FITC and propidium iodide (PI). After treatment with 10 μM 5 for 48 h, SJSA-1 cells were labeled with Annexin-V and PI and analyzed by flow cytometry. Representative results of three independent experiments are shown. Other results are shown in Fig. S31–S33.† (c) Apoptotic responses of SJSA-1 cells (left) and p53-mutated SW480 cells (right) to 1, 5, 5-Rv, and Nutlin-3a. Error bars represent standard deviations of three independent experiments. *p < 0.05 and **p < 0.01 from t test between each sample and the vehicle control.