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. 2008 Jul 3;295(3):G512–G521. doi: 10.1152/ajpgi.90327.2008

Fig. 4.

Fig. 4.

A: PMA treatment overnight induces macrophage marker genes (CD14 and CD68) in peripheral blood monocytes (PBMs) isolated from healthy volunteers as determined by real-time PCR. PBMs were cultured overnight with the indicated concentrations of PMA and RNA extracted for CD14 and CD68 real-time PCR. *P < 0.05 compared with vehicle-treated cells. B: CD68-positive macrophages increase in number with PMA treatment at 100 and 200 nM. PBMs treated with PMA as above were analyzed for CD68 expression by fluorescence-activated cell sorting (FACS). *P < 0.05 compared with vehicle-treated cells. C: in both PMA-induced macrophages and vehicle (DMSO)-treated monocytes, the ILI response was determined by assessment of intracellular low molecular weight-Fe59 complexes ([LMW-Fe]i) in response to peroxynitrite (10 μM) following Fe59Cl3 labeling. Note PMA-induced macrophages but not monocytes exhibit the ILI response. D: peroxynitrite-induced TNF-α release by PMA-induced macrophages derived from PBMs is fourfold greater than vehicle-treated PBMs. L1 treatment abrogates this differentiation-associated, maximal TNF-α expression. *P < 0.05 compared with vehicle-treated PBMs (no PMA). +P < 0.05 compared with PMA-treated PBMs without L1 treatment.