Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2013 Oct 1;27(19):2086–2098. doi: 10.1101/gad.224899.113

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2013 Ruffell et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

PMC Copyright notice

Figure 5. — CtsC expression by fibroblasts and immune cells mediates angiogenesis and growth of transplantable tumors. (A) Deficient tumor growth in mice lacking CtsC. PDSC5 clone 6 (PDSC5.6) tumor cells were injected subcutaneously (s.c.) into CtsC^−/− versus CtsC^+/− syngeneic FVB/n mice. n = 20 mice per group. Significance between CtsC^−/− versus CtsC^+/− hosts was determined by an unpaired t-test; (*) P < 0.05. (B) Angiogenesis was attenuated as early as 6 d following implantation of PDSC5.6 tumor cells in CtsC^−/− mice compared with CtsC^−/− cohorts. Blood vessels were evaluated by CD31 immunohistochemistry. Values represent the average of five high-power fields of view per mouse. n = 4–14 tumors per category. Significance was determined by an unpaired t-test; (**) P < 0.01; (***) P < 0.001. (C) PDSC5.6 cells alone (blue) or admixed with BMD CD45⁺ isolated from HPV16/CtsC^+/− (orange) or HPV16/CtsC^−/− (green) mice at 4 mo of age were injected s.c. into CtsC^+/− (circle) or CtsC^−/− (square) at a ratio of 3:1 (PDSC5:CD45⁺). n = 8–9 mice per category. Significance between PDSC5 cells in combination with CD45^+/− derived from HPV16/CtsC^+/− versus PDSC5 cells alone in CtsC^+/− hosts was determined by an unpaired t-test; (*) P < 0.05. (D) Mast cells derived from CtsC^+/− and CtsC^−/− bone marrow and treated for IgE-dependent FcεRI stimulation were evaluated for HUVEC migration using a Boyden chamber assay. n = 3–4 mice per group. Significance was determined by an unpaired t-test; (*) P < 0.05. (E) HPV16/CtsC^+/−-derived NAFs isolated at 4 mo of age are necessary but not sufficient to mediate tumor growth in CtsC^−/− mice. PDSC5.6 cells alone (blue) or admixed with HPV16/CtsC^+/−-derived (orange) or HPV16/CtsC^−/−-derived (green) NAFs were injected s.c. into CtsC^+/− (circle) or CtsC^−/− (square) at a ratio of 3:1 (PDSC5:NAF). n = 5–14 mice per group. Significance between PDSC5 cells in combination with NAFs derived from HPV16/CtsC^+/− versus PDSC5 cells alone in CtsC^+/− hosts was determined by an unpaired t-test; (*) P < 0.05. (F) HPV16/CtsC^+/−-derived and HPV16/CtsC^+/−-derived NAFs at 1 and 4 mo of age were evaluated for HUVEC migration using a Boyden chamber assay. n = 3–5 mice per group; samples were assayed in triplicates. Significance was determined by an unpaired t-test; (*) P < 0.05. (G) Reconstitution of CtsC-deficient mice with CtsC^+/− bone marrow in combination with HPV16/CtsC^+/−-derived NAFs isolated at 4 mo of age was sufficient to restore and sustain tumor growth in the absence of host-derived CtsC. PDSC5.6 cells were admixed with HPV16/CtsC^+/−-derived or HPV16/CtsC^−/−-derived NAFs (ratio 3:1) and injected s.c. into CtsC^+/− (blue) or CtsC^−/− (red) host mice that were lethally irradiated and transplanted with bone marrow from CtsC^+/− or CtsC^−/− mice that also expressed GFP under control of the β-actin promoter. Shown are tumor volumes at day 55 after tumor cell inoculation. n = 3–5 mice per group. Significance was determined by an unpaired t-test; (*) P < 0.05; (**) P < 0.01. (H) Density of angiogenic vasculature from G was evaluated by CD31 immunohistochemistry. Values represent the average of five high-power fields of view per mouse. n = 3–5 tumors per category. Significance was determined by an unpaired t-test; (*) P < 0.05; (***) P < 0.001.