Sigmaplot 12.0 for windows

Inhibition curves were fitted using the non-linear regression [Inhibitor] vs. response – Variable slope (four parameters) model on GraphPad Prism 7.03 (GraphPad Software). Maximum inhibition and half maximal inhibitory concentration (IC50) values were calculated for each experimental repeat and reported as mean ± SD (n ≥ 3). Statistical analyses were performed with the two-tailed unpaired t-test. P < 0.05 was considered statistically significant.
Stopped-flow fluorescence traces were analyzed using a Pro-Data Viewer (Applied Photophysics Ltd). Assuming pseudo-first-order kinetics with [PA] at least 5-fold higher than that of NBD P9 PAI-1 or its complex with Nbs, a single exponential equation was fit to the stopped flow traces to calculate k_obs. The quality of the fit was estimated by visual analysis of the plots of the residuals (deviation of the fitted function from the actual data). The values of k_obs (n ≥ 3; standard error < 10%) were plotted against [PA]. Plots were fitted by hyperbolic or linear equation to calculate parameters (k_lim, Km, and k_lim/Km), using nonlinear least squares fitting with the Levenberg-Marquardt algorithm (SigmaPlot 12.0 for Windows; SPSS Inc). Correlation coefficients (r) calculated from curve fittings were used as a parameter of the goodness of fit; r² of the fit was greater than 0.90 for all the kinetic data.

The following dependent variables were analyzed using a two-tailed independent Student's t-test (open-field, social interaction, and 5-HT, norepinephrine, and dopamine concentrations). Fear conditioned freezing behavior was analyzed using a one way ANOVA with repeated measures with drug treatment as main factor and time as the repeated measures. In the presence of significant main effects, between-subjects post-hoc tests were conducted using two-tailed independent Student's t-tests. Statistical significance was accepted with p ≤ 0.05. All statistical analyses were carried out using SPSS 22.0 (SPSS Inc., Chicago, IL, USA) and all graphs were generated using SigmaPlot 12.0 for Windows (SPSS Inc.) and figure-plate illustrations were done using CorelDraw version 12 for Windows.