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. 2006 Aug 24;36(1):68–77. doi: 10.1165/rcmb.2006-0165OC

Figure 2.

Figure 2.

Inhibition of endogenous ROS by antioxidants reduces cell survival, phosphorylation of Akt and p38 MAPK in human monocytes. (A) Monocytes (1 × 106/condition) were isolated and incubated with DPI (10 μM), NAC (20 mM), or DMSO control for 1 h, then stimulated with (+) or without (−) M-CSF for an additional 24 h and stained with Annexin V/PI for flow cytometry. Unstimulated monocytes were used as the 100% Annexin V/PI control. Data are expressed as the mean ± SEM of six independent donors (*P < 0.01 compared with M-CSF alone control). (B) Monocytes (10 × 106/ condition) pretreated for 1 h with the indicated inhibitors were either unstimulated (−) or stimulated (+) with M-CSF (100 ng/ml) for 6 h, then lysed. An equal amount of protein was subjected to Western blot analysis using antibodies recognizing pAktThr308 and pAktSer473 or phospho-p38Thr180/Tyr182. (C) The membranes were stripped and reprobed for Akt1 or p38 MAPK. Shown in B and C is a representative blot from six independent donors.