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. 2014 Nov 19;34(12):2554–2562. doi: 10.1161/ATVBAHA.114.304717

Figure 4.

Figure 4.

Serum-starvation induces production of reactive oxygen species, which is blocked by CX3C chemokine fractalkine (CX3CL1) in classical monocytes. A, Monocytes were loaded with CellROX green reagent, and then incubated for 30 minutes with serum-free medium (SFM), fetal calf serum (FCS), CX3CL1, or CCL2 then analyzed by flow cytometry. Data are shown as geometric mean of 3 to 12 donors from 2 to 6 experiments. B, Cells were treated as in A with SFM, FCS, or full-length (FL) CX3CL1. n=4 donors from 2 experiments. C, Monocytes were loaded with dihydroethidium then treated with agonists as in A. Levels of 2-hydroxyethidium were analyzed by high performance liquid chromatography (HPLC), data shown as fold change relative to FCS sample for each donor, n=4 donors from 2 experiments. D, Cells were loaded with CellROX green±2 μmol/L AZ12201182 (AZ) then incubated with SFM, FCS, or CX3CL1. n=8 donors from 4 experiments. E, Classical and nonclassical monocytes were isolated from the same donor as described in Figure 2 then treated as in A. Data shown as fold change relative to FCS sample for each donor. n=5 donors from 2 experiments. Data shown as mean+SEM. Data were analyzed by 1-way ANOVA and Dunnett (A–D) or Sidak (E) post hoc test, *P<0.05, **P<0.01, ***P<0.001, ns=not significant relative to SFM.