Figure 1
AEZS-136 treatment affects the PI3K/AKT and MAPK pathways, reduces cell proliferation, and increases cell death.
(a) Immunoblots of extracts from HL cells treated with AEZS-136 (10 μM) or DMSO vehicle for 0.5, 1, or 2 hours. Equal protein loading was confirmed by immunoblotting for β-actin. The experiments were repeated twice with similar results. Representative blots are shown. (b) Dose-dependent effect of AEZS-136 (1.25–20 μM) on phosphorylated and total AKT and ERK after exposure to AEZS-136 or DMSO vehicle for 2 hours in HL cells. (c) HL cells were exposed to 2.5 μM (green), 5 μM (blue), or 10 μM (red) AEZS-136 for 24, 48, or 72 hours, after which, the cell proliferation was assessed as described in the Methods section. The mean (±SEM) values refer to three independent experiments. *P ≤ 0.0001, ≠P ≤ 0.001, °P ≤ 0.01, and ≈P ≤ 0.05 compared with vehicle-treated cells. (d) Cell cycle analysis of AEZS-136 (10 μM)-treated L-540, SUP-HD1, KM-H2, and L-428 cells or vehicle-treated controls after 48 hours. Flow cytometry was performed to measure the cell cycle distribution of treated cells compared with untreated controls. The mean (±SEM) values refer to three independent experiments. *P ≤ 0.05 and **P ≤ 0.01 compared with vehicle-treated cells. (e) Immunoblots of extracts from HL cells treated with AEZS-136 (10 μM) or vehicle-treated controls for 48 hours. Equal protein loading was confirmed by immunoblotting for β-actin. The experiments were repeated twice with similar results. Representative blots are shown. (f) HL cells were exposed to 2.5 μM (green), 5 μM (blue), and 10 μM (red) AEZS-136 for 24, 48, and 72 hours, after which, the percentage of dead cells (Annexin V+/PI+ plus Annexin V−/PI+) was obtained as described in the Methods section. The mean (±SEM) values correspond to three independent experiments. *P ≤ 0.0001, ≠P ≤ 0.001, °P ≤ 0.01, and ≈P ≤ 0.05 compared with the vehicle-treated control cells.
