Figure 5.

A 15-LOX-1 inhibitor does not modify cytoprotection against UOS mediated by 32:6n3 and 34:6n3 on primary human RPE cells. Serum-deprived (a,b) and low serum (c,d) primary human RPE cells were incubated with the 15-lipoxygenase-1 (15-LOX-1) inhibitor (10 micromolar, PD146176) for 1 hour, then subjected to oxidative stress (H₂O₂/TNFα) for 16 hours to induce apoptosis (a–d,f). The addition of 32:6n3 and 34:6n3 protected human RPE cells (b,d,f) from cell death. Typical fields of cell cultures are represented in the right column. Nuclei are labeled with Hoechst staining, and the dead cells are highlighted in green. These were separated using an intensity threshold algorithm and counted using an Image J macro (left column)⁷⁰. (e) Quantification of live (control cells; black curve) and dead (UOS cells, red curve) cells was based on nuclear size⁷⁰. Error bars, SEM; *p < 0.05.