Uncoupling Protein 2 Regulates Palmitic Acid-Induced Hepatoma Cell Autophagy
Figure 1
PA induces autophagy in H4IIE cells. (a) H4IIE cells were treated with PA (6 h) conjugated to fatty acid-free BSA at different concentrations, or H4IIE cells were treated with PA (250 μM) conjugated to fatty acid-free BSA at different time points. H4IIE cells treated with BSA acted as a control. After treatments, cells were stained and subjected to the WST-1 assay. (b) Intracellular lipid accumulation was assessed with Nile red staining. PA-induced autophagy (250 μM) of H4IIE cells exhibited numerous small discrete bodies distributed throughout the cytoplasm (objective lens, ×40). (c) H4IIE cells were treated with PA (250 μM) conjugated to fatty acid-free BSA for 2, 4, 6, 8, 12, and 24 h as indicated. Cells treated with BSA acted as a control. After the treatment, cell lysates were collected and subjected to western blotting. (d) H4IIE cells were treated with BSA, PA (250 μM), or PA + CQ (10 μM) for 6 hours before being processed; then electron microscope was performed at 40,000x magnification. All values are the means ± SD of three independent experiments each performed in triplicate.
