|
|
Blood, Vol. 110, Issue 1, 142-150, July 1, 2007
CD13/APN regulates endothelial invasion and filopodia formation
Blood Petrovic et al.
110: 142
Supplemental materials for: Petrovic et al, Vol 110, Issue 1, 142-150
Files in this Data Supplement:
- Figure S1. Murine hemangioendothelioma-derived endothelial cell line (JPG, 37.6 KB)
-
A murine hemangioendothelioma-derived endothelial cell line (EOMA, which we have previously shown to express very low levels of endogenous CD13) was stably transfected with the CD13 expression plasmid (pcDNA-CD13), and 6 independent clones were isolated and characterized for their ability to bind and internalize bradykinin. The relative increase in cell-surface CD13 expression detected in the individual clones by FACS analysis was 5.08 ± 2.32 fold (data not shown). CD13-expressing cells showed a significant increase in relative bradykinin internalization (2.17 ± 0.81 fold, P = .024 using the Mann-Whitney nonparametric Student t-test) while maximum receptor binding of H3BK increased 1.60 ± 0.65 fold without reaching significance (P = .262), consistent with our findings using CD13 inhibitors.
- Figure S2. Filopodia contains actin (88.6 KB)
-
Cells were treated as in “Materials and methods” and stained with Texas Red phalloidin. Images were inverted using Adobe Photoshop.
- Figure S3. Structures visualized by filipin staining contain actin (JPG, 47.9 KB)
-
Overlays using Adobe Photoshop. Areas staining both red and blue (ie, both actin and cholesterol) become pink in overlay.
- Figure S4. Overlays of filipin/phalloidin double-stained HUVECs (JPG, 57.0 KB)
-
- Figure S5. Bestatin treatment does not affect total cholesterol levels (JPG, 47.7 KB)
-
Total cholesterol was measured in HUVECs using the Amplex Red Cholesterol Assay Kit (Invitrogen). (A) HUVECs were either untreated, treated with inhibitory concentrations of bestatin, or treated with inhibitory MY-7 antibodies in the presence or absence of bradykinin. (B) HUVECs were depleted of cholesterol with MBCD as described in “Materials and methods” (MBCD) and cholesterol was replaced in the presence or absence of bestatin.
- Figure S6. HUVECs were grown on Alcian Blue–coated coverslips with or without bestatin treatment and imaged as described in “Materials and methods” (JPG, 90.9 KB)
-
- Figure S7. Distribution of other light fraction resident proteins is affected by bestatin treatment (JPG, 50.9 KB)
-
|
|
