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. 2016 Jul 1;6:28811. doi: 10.1038/srep28811

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Copyright © 2016, Macmillan Publishers Limited

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(a) Motility of transfected JEG-3 cells in response to I-BOP (200 nM) was investigated in a chemokinesis assay over 48 hours and compared to vehicle control. The distance the cells moved was analysed using TScratch software and expressed as a percentage wound closure, n = 6. (b) Adhesion of transfected JEG-3 cells, pre-treated with 200 nM I-BOP for 24 hours, to different extracellular matrix proteins (collagen I, □; collagen IV, ; fibronectin, ; vitronectin, ■) was assessed over a 30 minute period and compared to vehicle control. Attachment was quantified using methylene blue staining. (c) Transfected JEG-3 cells, treated with vehicle or 200 nM I-BOP, were stained with anti-vinculin antibodies and imaged at 400x magnification via fluorescence microscopy to identify focal adhesions. Scale bar: 20 μm. (d) Changes in FAK activation/phosphorylation at specific residues was measured via immunoblotting in transfected JEG-3 cells treated with vehicle or 200nM I-BOP. Loading was determined by re-probing the membranes with non-phosphorylated FAK and protein bands were quantified using densitometry. The data are represented as mean ± SEM and are representative of 3 independent experiments. One-way ANOVA, NS: not significant, *P < 0.05, **P ≤ 0.01, ****P < 0.001.

Inline graphic — (a) Motility of transfected JEG-3 cells in response to I-BOP (200 nM) was investigated in a chemokinesis assay over 48 hours and compared to vehicle control. The distance the cells moved was analysed using TScratch software and expressed as a percentage wound closure, n = 6. (b) Adhesion of transfected JEG-3 cells, pre-treated with 200 nM I-BOP for 24 hours, to different extracellular matrix proteins (collagen I, □; collagen IV, ; fibronectin, ; vitronectin, ■) was assessed over a 30 minute period and compared to vehicle control. Attachment was quantified using methylene blue staining. (c) Transfected JEG-3 cells, treated with vehicle or 200 nM I-BOP, were stained with anti-vinculin antibodies and imaged at 400x magnification via fluorescence microscopy to identify focal adhesions. Scale bar: 20 μm. (d) Changes in FAK activation/phosphorylation at specific residues was measured via immunoblotting in transfected JEG-3 cells treated with vehicle or 200nM I-BOP. Loading was determined by re-probing the membranes with non-phosphorylated FAK and protein bands were quantified using densitometry. The data are represented as mean ± SEM and are representative of 3 independent experiments. One-way ANOVA, NS: not significant, *P < 0.05, **P ≤ 0.01, ****P < 0.001.