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Figure 1 | Particle and Fibre Toxicology

Figure 1

From: Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation

Figure 1

PM induces ROS-dependent EC barrier disruption and ZO-1 degradation. (A) Human lung microvascular ECs grown on 6-well plates were challenged with PM (100 μg/ml) for 0.5-6 hr. Cell lysates were analyzed by Western blotting with antibodies to ZO-1, ZO-2, VE-cadherin, and β-catenin. (B) Human lung microvascular ECs grown on 6-well plates were challenged with PM (100 μg/ml) for 1 or 6 hr with or without NAC (5 mM, 1 hr pretreatment). Cell lysates were analyzed by Western blotting. Changes in levels of ZO-1 are expressed as fold changes and normalized to control. Shown are representative blots from three independent experiments. Relative levels of ZO-1 densitometry are shown in bar graph under corresponding experimental conditions. N = 3. *p < 0.05 compared to control. **p < 0.05 compared to PM-6 hr group. (C) Human lung microvascular ECs grown on glass chamber slides were challenged with PM (100 μg/ml) for 6 h with or without NAC (5 mM, 1 hr pretreatment). VE-cadherin and ZO-1 localization was detected by immunofluorescence staining with antibodies to VE-cadherin and ZO-1. (D) ECs grown on ECIS gold electrodes were pretreated with NAC (5 mM, 1 hr) or PEG-CAT (250 U/ml, 1 hr), then challenged with PM at 100 μg/ml. Changes in TER were measured with the ECIS 6 hr post-PM treatment. *p < 0.05 compared to PM only group.

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