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. 2014 Apr 10;20(11):1693–1708. doi: 10.1089/ars.2013.5219

FIG. 3.

FIG. 3.

PACs lacking Nrf2 reveal lower expression of antioxidant genes and higher mortality under oxidative stress conditions. Mononuclear cells were isolated from bone marrow of Nrf2+/+ and Nrf2−/− mice and cultured for 7–10 days in vitro. (A) Analysis of surface marker expression. Flow cytometry (nNrf2+/+=3, nNrf2−/−=3); Nrf2 deficiency in PACs affects the mRNA level of (B) HO-1 and (C) NQO1. PACs were cultured for 24 h under normoxia or hypoxia (0.5% O2). RT-PCR (nNrf2+/+=3, nNrf2−/−=3). (D, E) PAC survival under conditions of oxidative stress is impaired in the absence of Nrf2. (D) LDH release after stimulation with H2O2 for 24 h (nNrf2+/+=3–9, nNrf2−/−=2–8). (E) The percentage of apoptotic/necrotic cells after stimulation with 200 μM H2O2 for 24 h. Staining, respectively, for annexin V (AnV) and propidium iodide (PI). Flow cytometry (nNrf2+/+=6, nNrf2−/−=6); each bar represents the mean±SEM. *p<0.05, control versus H2O2; #p<0.05, Nrf2+/+ versus Nrf2−/−. PACs, proangiogenic cells.