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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: FEBS J. 2016 Oct 1;283(21):3898–3918. doi: 10.1111/febs.13895

Figure 4.

Figure 4

Gal/VNPT55 decreases mesenchymal factors and enhances epithelial marker expression. (a) Lysates from PC-3 and DU145 cells treated with 5 and 10 μM were subjected to immunoblot analysis to evaluate N-Cadherin and ZO-1. (b) PC-3 and DU145 cells incubated with gal and VNPT55 at 10 μM for 24 h were stained with E-Cadherin polyclonal antibodies following protocol in materials and methods. Images were taken with Zeiss camera mounted immunofluorescence microscope. (c) PC-3, DU145 and CWR22Rv1 cells were analyzed for MMP-2/-9 protein expression after gal/VNPT55 treatment. (d) LNCaP cells were serum starved for 12 h and treated with 5, 10 and 20 μM gal/VNPT55 for 36 h in serum-free, pen-strep free RPMI media. Culture media was concentrated using Millipore 0.5 ml ultra-centrifugal columns. Conditioned media was separated on 0.1% gelatin Tris/Glycine gel to analyze proteolytic activity of MMPs (top), densitometric analyses (bottom bar chart) shows significant decrease in MMP-2 activity (*p<0.5, **p<0.001). (e) PC-3 cells were treated with gal/VNPT55 at 2.5 and 5 μM for 72 h and media concentrated as in (d) and a zymogram gel used to determine proteolytic activity of MMP-2/-9. (f) DU145 cells were treated as in (e), at 2.5 μM with gal/VNPT55 and analyzed with gelatin gels. Densitometric analysis shows a significant decrease in MMP-2/-9 activity. All zymogram assays were repeated at least three times and represented as means ± S.E.M (*p<0.05).