Fig. 4.

PC protection is blocked by 15-LOX-1 inhibition and is selectively rescued by exogenous NPD1. APRE-19 cells were treated with vehicle, PD-146176, NPD1, 15-HETE, or PEDF prior to mild oxidative preconditioning and lethal OS. Cell and media were collected for lipid analysis, and nuclei were imaged for cell survival using MAIM. a Experimental design. b Typical mass spectrometry chromatograms of 15-HETE, 17-HDHA, and NPD1 during PC with and without the 15-LOX-1 inhibitor PD-146176. c UPLC-ESI-MS/MS graphical quantification of 15-HETE, 17-HDHA, and NPD1 during PC with and without PD-146176. d, e Quantification of percentage of non-pyknotic nuclei challenged with lower concentrations of uncompensated OS with and without PD-146176 (2 µM) or PEDF (1 nM) with and without PD-146176 for 6 h prior to lethal challenge. f Representative 8-bit 10× zoomed-in micrographs of ARPE cells treated ± PC ± PD-146176 in combination with 100 nM 15-HETE or 100 nM NPD1. g Graphical quantification of percentage of non-pyknotic nuclei during lethal oxidative challenge with and without PC, PD-146176, NPD1, or 15-HETE. *p < 0.05, ^# p < 0.001, scale bar 10 µM