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. 2010 Jul 15;10(2):198–208. doi: 10.4161/cbt.10.2.12193

Figure 1.

Figure 1

Wnt1 transformation upregulates MMP-3 expression in C57MG cells and induces β-catenin translocation. (A) Conditioned media from C57MG and C57MG/Wnt1 clones (25 µg/lane) was loaded on gelatin (a), casein (b) and reverse gelatin zymographies (c). Conditioned media from NIH3T3 and HT1080 cells were used as positive controls for mouse MMP-2 and human MMP-2 and MMP-9 expression, respectively. (B) Gel was incubated overnight in the presence of AG3340 (20 µg/ml) (d). Samples were incubated with AP MA(10 µM) prior to electrophoresis (e). Recombinant mouse MMP-3 was loaded in one lane as positive control (f). (C) Western blot analysis of conditioned media (20 µg/lane) as in (A), of conditioned media from C57MG cells treated with Wnt3a, and of conditioned media and cell lysates of L and L/Wnt3a cells (g). Recombinant human MMP-7 and limb extracts from 15-day-old mouse embryos were used as positive controls for MMP-7, MMP-13 and MMP-14 respectively (h). (D) Photomicrographs of C57MG, C57MG/Wnt1 and C57MG cells treated with Wnt3a (top). Immunolocalization of β-catenin detected in the cell membrane (arrows) and in the nuclei (arrowheads) (bottom). Bar = 100 µm in phase contrast and 20 µm in fluorescence.