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. 2015 Jul;185(7):2000–2011. doi: 10.1016/j.ajpath.2015.03.022

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© 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

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Real-time quantitative PCR (qPCR) and immunoblot comparisons of Col5a2 expression levels. A: qPCR of RNA from Col5a2^+/+ and Col5a2^−/− mouse embryonic fibroblasts (MEFs) consistently revealed essentially undetectable levels of Col5a2 mRNA transcript in Col5a2^−/− MEFs, exhibiting large quantification cycle values outside the linear range of the primers. B: Immunoblotting was unable to detect α2(V) chains in the conditioned medium of Col5a2^−/− MEFs. C: qPCR reveals Col5a2 RNA levels of Col5a2^+/− dermal fibroblasts to be half that of Col5a2^+/+ dermal fibroblasts. D: Immunoblotting shows pro-α2(V) chain levels to be similarly reduced in the conditioned media of Col5a2^+/− compared with levels found in conditioned Col5a2^+/+ dermal fibroblast medium. ^∗∗P < 0.01.