Uncoupling Protein 2 Regulates Palmitic Acid-Induced Hepatoma Cell Autophagy
Figure 4
Inhibition of UCP2 decreases PA-mediated autophagy. H4IIE cells were transfected with UCP2-siRNA for 72 h to inhibit the expression of UCP2, followed by 250 μM PA for 6 h, with or without CQ (10 μM). (a) The level of UCP2 mRNA was normalized to 18 s, and UCP2 protein was normalized to -actin. This ratio was set to 100% in the control of BSA. (b) The level of LC3 protein was normalized to -actin, and this ratio was set to 100% in the control of BSA. (c) H4IIE cells were transfected with UCP2 siRNA for 72 h, followed by 250 μM PA for 6 h. Then LC3 puncta formation was observed using an inverted fluorescence microscope. The numbers of LC3 puncta/cell were counted from ≥100 cells. (d) Cells were treated with PA (250 μM) for 6 h, with or without CQ before being processed; then electron microscope was performed at 40,000x magnification. Data are expressed as the mean ± SD for each experiment. All data presented are representative of three separate experiments with consistent results.
