Display Settings:

Items per page

Results: 6

2.
Figure 5

Figure 5. Effects of CPI on angiogenesis in the Matrigel assay. From: VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation & mother vessel formation.

(A) Top panel is stained with H&E, bottom panel with antibodies to CD31. Dashed line indicates border between Matrigel and surrounding host tissue. Note prominent MV in Matrigels containing bFGF alone, and dearth of vessels in Ctl (no additions) and in bFGF Matrigels supplemented with stefin A, cystatin C or CA-074Me. Scale bar, 50 μm. (B) Microvascular density (mean ± SEM). *, p<0.01 all other conditions versus +F (+FGF), Dunn's multiple comparison test.

Sung-Hee Chang, et al. Cancer Res. ;69(10):4537-4544.
3.
Figure 6

Figure 6. Angiogenic response and vascular permeability in wild type and cathepsin B null mice 4 days after injection of Ad-VEGF-A164. From: VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation & mother vessel formation.

(A) Histology. Arrows identify some of the many MV present in wild type mice, whereas very few MV were found in null mice. Scale bar, 50 μm.
(B) Microvessel density (mean ± SEM) was calculated as described in Methods and analyzed statistically with an unpaired t test. *, p<0.0001.
(C) Evan's blue dye accumulation (mean ± SEM) was quantified using IPLab software. WT, n=5; CatB null, n=6. *, p=0.01, unpaired t test.

Sung-Hee Chang, et al. Cancer Res. ;69(10):4537-4544.
4.
Figure 3

Figure 3. CPI expression in Ad-VEGF-A164 injected ears. From: VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation & mother vessel formation.

(A, B) Quantitative RT-PCR demonstrating CPI mRNA expression patterns at indicated times after Ad-VEGFA164 injection (mean ± SD). Representative data from 9 different experiments performed on 7 different animals.
(C) Immunohistochemical staining of CPIs in control ears (time 0) and at 3 and 5 days after Ad-VEGF-A164 injection. Staining for all three CPIs is greatly reduced in mother vessels (MV) compared with normal venules (V). Arteriole (A) staining for cystatin C, but not for cystatin B or stefin A, was also reduced following Ad-VEGF-A164 injection. Scale bar, 50μm.

Sung-Hee Chang, et al. Cancer Res. ;69(10):4537-4544.
5.
Figure 1

Figure 1. Degradation of vascular basement membranes in MV induced by Ad-VEGF-A164 or by TA3/St tumors. From: VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation & mother vessel formation.

(A) Confocal microscopy of ear whole mounts from normal control (Ctl.) mice and from mice whose ears had been injected 3 days earlier with Ad-VEGF-A164. Immediately prior to sacrifice, mice were injected i.v. with FITC-lectin (green). Ears were then immunostained with antibodies against laminin or collagen IV and visualized with Texas Red conjugated secondary antibodies. Note extensive loss of red staining (laminin or collagen) in MV as compared with control venules. Scale bar, 50μm.
(B) Immunohistochemistry for laminin and collagen IV in mother vessels (MV) of TA3/St tumors harvested 4 days after implantation. Note patchily reduced MV staining (red arrowheads), compared with strong, continuous basement membrane staining of normal venules (insets). Scale bar, 50μm.
(C) Immunoblot with an antibody against laminin β1 chain performed on extracts of normal ears (time 0) and on ears harvested at 1-21 days following Ad-VEGF-A164 injection. Note increasing low molecular weight laminin β1 chain fragments (bracket) at 1-5 days. In contrast, on days 7 and 10 there is increased expression of intact laminin β1 chain (arrow), as well as high molecular weight fragments.
(D) Glomeruloid microvascular proliferations in ears 7 days after Ad-VEGF-A164 injection demonstrate extensive new laminin deposition (brown stain). Scale bar, 50μm.

Sung-Hee Chang, et al. Cancer Res. ;69(10):4537-4544.
6.
Figure 2

Figure 2. Cathepsin expression in developing MV. From: VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation & mother vessel formation.

(A) Quantitative RT-PCR demonstrating expression patterns (mean ± SD) of cathepsin B, D, L, H and S mRNAs at indicated times after Ad-VEGF-A164 injection. Data are representative of nine different experiments performed on 7 different animals.
(B) Immunohistochemical localization of cathepsin B in control ears (time 0) and at 1, 3 and 5 days after Ad-VEGF-A164 injection. Cathepsin B is expressed by many different cell types in normal ears but increased expression following Ad-VEGF-A164 injection was observed only in mother vessels (MV). V, normal venule; A, arteriole. Scale bar, 50μm.
(C, top panel) Hydrolysis of Z-Arg-Arg-AMC (mean ± SD), a synthetic substrate commonly used to measure cathepsin B activity, in extracts of mouse ears that had been injected with Ad-VEGF-A164 or Ad-LacZ. Data are representative of 6 separate experiments. (C, middle panel) Fluorescence scan demonstrates cathepsin activity in extracts of mouse ears injected i.v. with GB123 at indicated times after Ad-VEGF-A164 or Ad-LacZ injection. Bands at 32 kDa, 28kDa, and 24 kDa were identified as cathepsins B, S and L, respectively. (C, bottom panel) Densitometry of pooled gel data (mean ± SD).
(D) Confocal microscopy of GB123 (red) and FITC-lectin (green) in normal mouse ears (NM) and in ears injected 3 and 5 days previously with Ad-VEGF-A164. Only rare stromal cells stain for GB123 (red) in NM ears, whereas perivascular cells, but not endothelial cells (white arrows), stain intensely in Ad-VEGF-A164-injected ears. White * indicate detaching GB123-positive perivenular cells. Blue color, DAPI staining. Scale bar, 20 μm.

Sung-Hee Chang, et al. Cancer Res. ;69(10):4537-4544.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk