Research Paper Volume 12, Issue 14 pp 13958—13978

Decreased autophagy and fuel switching occur in a senescent hepatic cell model system


Figure 1. Senescence induction in mice normal hepatic cells AML12. (A) Schematic representation of experimental strategy for the senescence induction. 4x106 Cells were seeded at day 0 in the T175 cell culture flask. Next day (day 1) cells were treated with 1.0 mM H2O2 followed by 750 μM for the subsequent 5 days. Cells can be sub-cultured at 1:3 ratio on day 3 if required. At day 7, cells were re-seeded for experiments as required. (B) Visualization of senescence induction in mice AML12 cells during H2O2 treatments from day 3, day 5, and day 7 before re-seeding. The images are taken randomly at 10x magnification. Scale bars as 100 μm. (C) 0.3x106 control and senescent AML12 cells were seeded in each well of the 6-well plate. Cells were trypsinized at indicated days and counted by an automated cell counter. (D and E) RT-qPCR analysis of senescence genes in AML12 cells (D) and liver tissues from young and old mice (E). * Statistical differences were calculated significant as p<0.05.