Figure 3.

Protective effect of D-limonene against H₂O₂–induced apoptosis. HLECs were pretreated with 1000 μM D-limonene for 12 h and then treated with 100 μM H₂O₂ for 24 h. The cells were then stained with Hoechst 33342 and observed using a fluorescence microscope. (a) Morphological apoptosis was observed using an inverted microscope. Cells that were exposed to H₂O₂ exhibited morphological changes typical of apoptosis, such as chromatin condensation and nuclear shrinkage. (b) The statistical analysis of the nuclear morphological change is shown. At least 70 cells in three different fields were counted per well in three separate experiments. ^∗∗ P < 0.01.