Functional confirmation of microarray data with Western Blotting and by Flow Cytometry – Presenting up-regulation of anti-apoptosis and down-regulation of pro-apoptosis proteins.

<p>(A) Western blotting quantification; 15 µg of protein from total cell lysate of PMN-B and PMN-O from chronic periodontitis patients were loaded, expression was normalized to β-actin used as internal control (n = 4). (B) Representative Western Blot of members of apoptosis pathway Bcl-xl (anti-apoptosis), Bim and Bax (pro-apoptosis). (C) Summary of relative changes in BCL-2 family (Ratio of expression in PMN-O compared to PMN-B). (D) Percentages of viable cells were analyzed by Flow cytometry in oral neutrophils (n = 3). Cells that were negative for Annexin V and PI were considered viable. * p ≤ 0.05 vs. Healthy. All data are mean ± SEM from 3 independent experiments.