Release of Matrix Metalloproteinases-2 and 9 by S-Nitrosylated Caveolin-1 Contributes to Degradation of Extracellular Matrix in tPA-Treated Hypoxic Endothelial Cells (original) (raw)

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Fig 3

OGD+tPA treatment (O+t) promotes Cav-1 secretion from bEND cells.

A) Upper panel: representative immunoblots showed that Cav-1 was detected in the conditioned media of tPA+OGD-treated cells (O+t), but not in untreated control cells (Ctrl) and the cells treated by OGD or tPA alone. Bottom panel: β-actin was not detected in the media of any cells. Experiments were repeated three times with similar results. B) Western blot analysis showed that OGD+tPA treatment (O+t) induced Cav-1 reduction in endothelial cells. Upper panel: representative immunoblots, β-actin served as loading control; Bottom panel: band intensity was quantitated after normalization to β-actin and the data were expressed as mean ± SD, *p < 0.05, ANOVA, N = 3. C) Confocal microscopic images revealed a circumcellular staining for Cav-1 protein in untreated control bEND3 cells, and OGD+tPA treatment (O+t) induced a significant reduction in Cav-1 immunostaining. Immunostaining experiments were repeated three times with similar results. Scale bar: 10 μm. D) Real time RT-PCR showed that OGD, tPA or their combination OGD+tPA (O+t) had no effect on Cav-1 mRNA expression level. Data were expressed as mean ± SD, ANOVA, N = 3.

Fig 3

doi: https://doi.org/10.1371/journal.pone.0149269.g003