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. 2015 Jan;94(1):12–21. doi: 10.1016/j.ejcb.2014.10.001

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© 2014 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

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Fig. 2 — Effect of TNF-α and GM6001 on type I collagen turnover assessed by the biochemical markers of degradation ICTP (A), neosynthesis CICP (B) and type I collagen formation (C and D). (A and B) Media from five explants per donor were pooled and then analyzed. CICP levels after ultrafiltration of day-4 samples are indicated by lower bars (B). Mean ± SEM. ^**p < 0.01, ^***p < 0.005 versus control at respective time point. (C and D) Western blot analysis for the α1 chain of type I collagen and β-actin of pooled concentrated (Amicon^® Ultra; Millipore) CNTZ tissue extracts (C and D) and conditioned media (C) from 30 individual 8-mm skin explants (5 explants from each of the 6 donors). (C) Loading of tissue extracts was normalized to the β-actin content determined separately (D) and media were adjusted to the corresponding volume to biopsy weight ratio. Lane 1, native skin; 2, control; 3, GM6001 (10 μM); 4, TNF-α. (D) Equal volume (12.5 μl) of the tissue extracts was applied to each well.