Fig. 4

Assay for protection against NMDA-induced mitochondrial membrane potential breakdown. a Three images from a TMRE time-lapse experiment, acquired at the indicated times after NMDA application. Colorization represents background-corrected TMRE intensity (see calibration bar). b Analysis of time lapse experiments. The upper left graph shows individual traces for untreated and NMDA-treated cells from four independent experiments (light colored traces depict the means of 3–10 replicate wells per experiment) and the mean from all experiments (bold traces). The remaining graphs show individual and mean traces for cells that were treated with compound + NMDA. The mean traces for untreated and NMDA-only treated cells are included in each graph for comparison. TMRE intensity was normalized to that of the first image which was taken immediately after NMDA application (time = 0). c Quantification of NMDA-induced loss of TMRE fluorescence. On the left graph, dots depict the mean of each individual experiment and vertical error bars indicate the SD of all experiments. On the right graph, mean differences are plotted as bootstrap sampling distributions. Each mean difference is depicted as a dot. Each 95% confidence interval is indicated by the vertical error bars. The p values of the two-sided permutation t-tests are: E, 0.0226; D, 0.006; O, 0.0006; F, 0.0376; I, 0.487; untreated, 0.006. For each permutation p value, 5000 reshuffles of the control and test labels were performed