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. Author manuscript; available in PMC: 2019 Sep 9.
Published in final edited form as: J Mol Cell Cardiol. 2018 Jul 5;121:107–123. doi: 10.1016/j.yjmcc.2018.07.003

Fig. 5.

Fig. 5.

TWEAK induces collagen, collagenase and gelatinase expression, and cardiac fibroblast proliferation and migration in part via TRAF3IP2. TWEAK (10 ng/ml) induced the expression of fibrillar collagens Iα1 and IIIα1 in a time-dependent manner (A), an effect blunted by TRAF3IP2 knockdown by lentiviral TRAF3IP2 shRNA (moi0.5 for 48 h) (B). Further, silencing TRAF3IP2 inhibited TWEAK-induced collagen secretion at 24 h (C; hydroxyproline assay), the collagenase MMP-13, MMP-14 and TIMP-1 mRNA (RT-qPCR) and protein expression (immunoblotting) at 2 h (D; protein expression is shown in insets at the top). Moreover, silencing TRAF3IP2 inhibited activation of the gelatinases MMPs 2 and 9 (E, F; immunoblotting using antibodies that detect both pro and active forms), as well as CF proliferation (CyQUANT® assay, G) and migration (Matrigel™ migration assay, H). A, B, D, E, A representative image from three independent immunoblot experiments with similar results is shown at the top and summarized semi-quantification of intensity of immunoreactive bands by densitometry from all three experiments in the respective right hand or bottom panels. C, G, H, n = 6. A-H, *P < at least 0.05 versus untreated, †P < 0.05 versus TWEAK, §P < 0.05 versus Untreated.