For calcium signal quantification, the number of individual peaks were difficult to quantify, since there were either large bursts of activity spanning tens of seconds (likely the result of trains of action potentials) or no network bursts seen prior to CNO addition. When there are no peaks prior to CNO addition, it is impossible to establish a ratio of firing based on peak number. Therefore, we used the area under curves (AUC) to quantify the degree of total network activity. To normalize all signals to the baseline, a custom-made MATLAB script containing both baseline subtraction by least-squares fitting was performed followed by intensity curve area calculation using the trapezoid rule. The MATLAB script prints an output of the intensity curve areas before and following CNO addition (highlighted in red and blue, respectively, Fig. 7A and B). Results were processed as (fluorescence curve area of post-CNO)/(area of pre-CNO) ratio, determined in the analysis time windows over all conditions (Fig. 7C). While there was variation in the area ratio between wells of the same condition, the CNO concentrations of 500 nM and
