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. 2016 Apr 22;11(4):e0153806. doi: 10.1371/journal.pone.0153806

Fig 5. The effect of high glucose culture on MMPs mRNA levels and proteolytic activity in primary human skin dermal fibroblasts.

Fig 5

Cell were cultured for 48 h in low (5mM) or high (25 mM) glucose media (see Methods for details). The effects of high glucose on the levels of MMPs mRNA expression were quantified by real-time RT-PCR. MMPs mRNA levels were normalized to the housekeeping gene 36B4, as an internal control for quantification. Primary human skin dermal fibroblasts do not express MMP-8 and MMP-20. Mean±SEM. N = 4. *p<0.05. Proteolytic activities from the conditioned media were examined by zymography, as described in “Methods”. Areas of protease activity will appear as clear bands against a dark blue background where the protease has digested the substrate. Data are representative of three experiments. MMPs inhibitor (GM60001) was used as specificity of MMPs-mediated proteolytic activity (last two lanes).