Study of cell migration and cytokine expression in pathogen infected oral keratinocytes.

<p><b>A.</b> Cell migration was assessed using transwell migration of keratinocytes after incubation with bacteria. The number of cells was counted by DAPI staining of nuclei using NIS element software under fluorescence microscope. Values were expressed as percent maximum against untreated control. Data are means ± SEM and shows one way ANOVA, with level of significance *p<0.05. <b>B.</b> Quantitation of integrin beta-3 at the protein level was determined by measuring fluorescence intensity of integrin beta-3 normalized to cell numbers. Values represent means ± SEM from three independent experiments. Significant between control and treatment *p<0.05 and between the two treatment groups +p<0.05 was determined by Ttest. <b>C–D.</b> Gene expression analysis of integrin beta-3 and -6 on day 1 and 3 post infection by real time RT PCR. The data is average from three independent experiments, δδCt gene/L32 value of each gene was calculated with normalization against expression of L32 control gene. Statistical significance represented by *p<0.05, and significant difference between expression on day 1 and 3 in a treatment group is represented by +p<0.05 after Tukey HSD test. <b>E–F</b>. mRNA levels of TNFα and IL-6 were measured by real time RT-PCR. Data shown are mean from three independent experiments. Statistical significance in differences of expression against control is represented by *p<0.05 after Ttest.