Figure 6.

LOBCE-induced migration and apoptosis protection are ROS-dependent. (A) PMNs (5 × 10⁴ cells per well) were incubated in the presence or absence of LOCBE (3 μg/mL) for one h. Then, PMNs were washed, incubated with luminol (100 μM), and total ROS production was measured by chemiluminescence emitted from luminol oxidation (n = 3). (B) PMNs were pre-incubated in the presence or absence of Trolox™ (100 μM) for 5 min before insertion in the upper compartment of a 48-well Boyden chamber (5 × 10⁴ cells per well), and LOBCE (3 μg/mL) was inserted in the lower compartment. DMEM medium alone was used as a negative control. PMNs migrated through a 5 μm pore-sized polyvinylpyrrolidone-free polycarbonate membrane. *** p < 0.001 compared to LOBCE group alone. (n = 3) (C) PMNs (10⁶ cells) were pre-incubated in the presence or absence of Trolox™ (100 μM) for 5 min. Then, PMNs were treated or not with LOCBE (3 μg/mL) for 20 h. After this time, PMN were centrifuged and stained with Diff-Quick™ staining kit, and apoptotic cells were counted using optical microscopy. *** p < 0.001 compared to the control group at 0 h, ^### p < 0.001 compared to the LOBCE 3 μg/mL + Trolox group (n = 3).