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gAd enhances the motility of mSAT.

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posted on 2012-05-18, 01:40 authored by Tania Fiaschi, Elisa Giannoni, Maria Letizia Taddei, Paola Chiarugi

A) Wound healing assay. Confluent mSAT were serum deprived overnight and then we performed an artificial wound. Cells were treated with free-serum medium (control), free-serum medium containing gAd (1 ug/ml) or 10% FCS. Cells were photographed after 5 hours. Black lines are indicative of the remaining gap in the wound. B) Migration of mSAT induced by gAd. mSAT were serum deprived overnight and then seeded in the upper chamber of the Boyden assay in free-serum medium with or without gAd (1 µg/ml). Four days differentiated C2C12 myotubes were cultivated in the lower chamber and used as chemo-attractant, *p<0,001 vs control. C) The invasiveness assay was performed as described in B, except for the presence of a thin layer of Matrigel in the upper chamber, *p<0,001 and ♦p<0,01 vs control. D) gAd enhances the production of MMP-2. mSAT were treated for 18 h with serum-free medium with or without gAd (1 µg/ml) and the corresponding media were subjected to zymography analysis. *p<0,001 vs control. E) The invasiveness assay of mSAT in response to gAd was performed as in C, except for the presence of the Ilomastat (broad MMP inhibitor) or aprotinin (uPA inhibitor), *p<0,001 and ♦p<0,01 vs control. Representative images of migrated mSAT, after staining with hematoxylin-eosin, are shown in B, C and E. Bar graphs represent the mean of migrated cells counted in six different fields for each experiment.

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