Fig. 1
Effect of organic matters from extracted ambient PM on VSMC migration. a Effects of seasonal organic matters from ambient PM on VSMC migration were measured using a wound healing assay (upper) and Boyden chamber assay (lower). The images from the wound healing assay are representative of five independent experiments, taken at the time of scratching and 48 h after scratching. The black line represents the initial boundaries following the scratch at time 0 and the white dotted line represents the migrating cell front after 48 h. The wound healing area was measured using ImageJ software. Images from the Boyden chamber assay shows the number of migrated cells. The images from the transwell assay are representative of five independent experiments, taken 12 h after seeding. b The effects of ambient PM on VSMC proliferation were evaluated using the BrdU incorporation assay. Serum-starved VSMCs were treated with negative control (0.1% DMSO), SPM, WPM or PDGF-BB (20 ng/mL) as a positive control for 24 h. c ROS generation in PM-treated VSMCs was evaluated using H2DCF-DA (green). VSMCs were pretreated with or without NAC (1 mM) for 1 h, then treated with negative control (0.1% DMSO), SPM or WPM. Nuclei were stained with DAPI (blue). Fluorescence intensity was quantified by SIBIA software. d Effects of the antioxidant, NAC, on organic matters from ambient PM-induced VSMC migration were measured using a wound healing assay (upper) and Boyden chamber assay (lower). All values are represented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 versus control; #P < 0.05, ##P < 0.01, and ###P < 0.001 WPM versus SPM; $P < 0.05, $$P < 0.01, and $$$P < 0.001 PM with NAC versus PM alone; NS, no significance