Figure 3.

Cadmium increases lung macrophage mitochondrial ROS. A, NF- κB activity was measured by luciferase assay in THP-1 cells exposed to vehicle, CdCl₂ (50 μm), IL-4 (20 ng/ml, negative control), or LPS (100 μg/ml, positive control) for 3 h (n = 6). Immunoblot analysis of exposed THP-1 cells in isolated (B) nuclear and (C) cytosolic extracts. mtROS generation in exposed THP-1 cells by (D) pHPA assay (n = 9) and (E) mitoSOX (antimycin A, 100 μm for 30 min, positive control) (n = 10). F, mtROS generation in BAL cells isolated from WT mice exposed to vehicle or CdCl₂ for 7 days (n = 6). Immunoblot analysis of BAL cells isolated from exposed mice in (G) nuclear and (H) cytosolic extracts. THP-1 cells were treated with vehicle or mitoTEMPO (10 μm, 16 h) and exposed to vehicle or CdCl₂ for 3 h. I, mtROS generation (n = 3). Immunoblot analysis in isolated (J) nuclear and (K) cytosolic fractions. L, TNFα; M, iNOS; N, arginase 1; O, TGF-β1; P, IL-10; and Q, PDGF-B mRNA expression (n = 3). ***, p < 0.0001. Values are shown as mean ± S.D.