Fig. 8
ZnT2 over-expression in MDA-MB-231 cells causes alterations in cell cycle, increased apoptosis and decreased invasion. a Cell cycle analysis was measured using in Mock-transfected and ZnT2 over-expressing (ZnT2-OX) MDA-MB-231 cells by flow cytometry. Data represent mean % of cells ± SEM, n = 3 samples/genotype from 3 independent experiments. Asterisk(s) indicates a significant difference from Mock transfected controls, *p < 0.05; **p < 0.01. b Cyclin dependent kinase 2 (CDK2) kinase activity in MDA-MB-231 cells. (Top panel) Representative immunoblot of immunoprecipitated CDK2, total CDK2 and β-actin in Mock-transfected and ZnT2-OX MDA-MB-231 cells. (Bottom panel) CDK2 kinase activity was assessed using histone H1 as a substrate. Representative gel illustrating 32P labeling of histone H1 and corresponding relative densitometric analysis. Data represent mean band intensity (normalized to CDK2 protein) ± SD, n = 3 samples/genotype, from 2 independent experiments. Asterisks indicates a significant difference from Mock transfected controls, ***p < 0.001. c Cell proliferation in Mock-transfected and ZnT2-OX MDA-MB-231 cells. Data represent mean absorbance ± SEM, n = 3 samples/genotype, from 3 independent experiments. Asterisk(s) indicates a significant difference from Mock-transfected controls, *p < 0.05; **p < 0.01. d Representative forward-by-side scatter plot depicts the population of early apoptotic (I), apoptotic (IV), apoptotic/dead (III) and dead cells (II). e The percentage of apoptotic and dead cells was significantly higher in ZnT2-OX cells compared to Mock-transfected cells. Data represent mean % cells ± SEM n = 3 samples/genotype from 2 independent experiments. Asterisks indicates a significant difference from Mock-transfected controls, ***p < 0.001. f ZnT2-OX resulted in a significant reduction in invasion (~80 %) compared with Mock-transfected controls. Data represent mean % control ± SEM n = 3 samples/genotype from 2 independent experiments. Asterisk indicates a significant difference from Mock-transfected controls, *p < 0.05