Figure 5

SAEC exposure to MWCNT induces ROS production and morphological changes in HMVEC. A. SAEC and HMVEC were grown in the apical and basolateral chamber, respectively, of a co-culture system and SAEC exposed to 1.2 μg/ml MWCNT for 6 or 24 h. ROS production (a-c) was assayed by the addition of 5 mM dihydroethidium to the basolateral media 30 min prior to fixation for confocal microscopy. Actin filaments (d-f) and VE-cadherin (g-i) were visualized by antibody staining. Red arrows show stress fibers, long white arrows show cellular gaps, and short white arrows show areas of actin ruffling. B. DHE fluorescence pixel intensity was quantified on the acquired confocal images using Optimas 6.51 software. A minimum of 10 cells were scored for each sample from 4 independent experiments. Data are presented as mean ± SE. There was a significant increase in average DHE pixel intensity at both 6 (23.62 ± 1.75) and 24 h (26.47 ± 1.06) over DM control cells (16.84 ± 1.45). * p < 0.05.