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Fig. 7 | BMC Biology

Fig. 7

From: The mitochondrially-localized nucleoside diphosphate kinase D (NME4) is a novel metastasis suppressor

Fig. 7

Migration, adhesion, and MMP activity of MDA-MB-231 cells genetically modified for NDPK-D. A Representative light microscopy images of MDA-MB-231 cell wound healing assay. Time 0 represents confluent monolayer wounds at 0 h and wounds were monitored 24 h after performing the scratch, in which empty vector control (CTR) monolayers became fully closed. Two different clones for each condition were studied. Images are representative of three independent biological replicates. Scale bar: 100 μm. B Representative light microscopy images of MDA-MB-231 dispase-based cell aggregation assay. Images are representative of three independent biological replicates; at least thirty pictures were analyzed for each replicate. Two different clones for each condition were studied. Scale bar 100 μm. C The size of the aggregates observed in B is depicted as the area of their horizontal projections. Data show means ± SEM of three independent biological replicates imaged. *****p< 0.00001 relative to control/empty vector (CTR). D Left panel, top, representative images of MMP activity by gelatin degradation zymography; the degradation bands of MMP9 and MMP2 are detected at 92 KDa and 72 KDa respectively. Left panel, bottom, representative Coomassie brilliant blue (CBB) of samples run simultaneously is shown as a loading control. Right panel, bar graphs represent the densitometric and statistical analyses of the bands obtained by gelatin zymography shown for MMP9 and MMP2 of four independent biological replicates. Concentrated culture media from MCF7 cells was used as positive control. Two different clones for each condition were studied. Data show means ± SEM (n=4). *****p< 0.00001 relative to control/empty vector (CTR). E Type I collagen invasion assay of MDA-MB-231 cells. Two different clones for each condition were studied. Data show means ± SEM. ***p< 0.001 relative to control/empty vector (CTR). Abbr. of MDA-MB-231 clones according to the expressed NDPK-D: CTR, control/empty vector; WT, wild-type; BD, CL-binding-deficient mutant; KD, kinase-dead mutant

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