Figure 2.

Neuroglobin (Ngb) overexpression combined with c-jun N-terminal kinase (JNK) inhibition protects against hypoxia/reoxygenation (H/R) injury in vitro. Normoxic control cells (solid bars) received the same treatment as cells exposed to 9 hours of hypoxia with 24 hours of reoxygenation (hypoxic; open bars). Oxidative stress assays for H/R injury: (A) reactive oxygen species (ROS) generation detected by electron paramagnetic resonance (EPR); (B) lipid peroxidation levels detected by malondialdehyde (MDA) assay; and apoptosis by (C) apoptotic cell death enzyme-linked immunosorbent assay (ELISA) and (D) caspase-3 (green) immunocytochemistry (ICC; nuclei=blue, 4',6-diamidino-2-phenylindole (DAPI). Scale bar, 100 μm. Data presented as mean±s.e.m., analyzed using unpaired Student's t-test and Bonferroni's post hoc correction, representative of n=3. *P<0.01 versus untreated hypoxic cells, ^†P<0.001 versus lenti-Ngb- or SP600125-treated hypoxic cells, ^#P<0.01 versus lenti-Ngb-treated hypoxic cells. DMSO, dimethyl sulfoxide; GFP, green fluorescent protein.