Figure 5.

High H₂O₂ level leads to escape from senescence: changed expression of proteins related to proliferation and EMT. Cells were subjected to 2.5 μM of IRINO and 5 or 10 μM H₂O₂. After 24 h, the medium was changed and the cells were cultured in a drug-free medium for 4 (■) or 7 (□) days. In a 7-day long variant of the experiment, after 4 days of cell culture, the medium was changed to the new one. (A) Production of protein related to cell cycle inhibition, geroconversion, and tumor suppression. Representative blots showing levels of p21, p-S6, and p53 proteins. (B) Production of proteins related to cell cycle progression. Representative blots show levels of cyclin A, cyclin B, p-cdc2, and p-Rb. (C) Production of antioxidative enzymes. Representative blots show levels of catalase, GPx-1, and HO-1 proteins. (D) Production of proteins related to EMT and stemness. Representative blots show levels of E-cadherin, Snail, and Nanog. GAPDH acts as a loading control.