| Description: |
A. Dynamics of nuclear enrichment of the dPSTR R under the control of the p AGA1 (dark blue), the p FIG1 (magenta) or the synthetic promoter with two PREs in tail-to-tail orientation with 3 bp spacing (p SYN 3TT - cyan). The solid line represents the median of the population, while the shaded area represents the 25- to 75-percentiles of the population. B. Dynamics of nuclear enrichment of the dPSTR Y under the control of the endogenous p AGA1 promoter which is present in parallel to the test dPSTR R reporter for the three strains presented in panel A. The response of the p AGA1 -dPSTR Y serves as a control for the robustness of pheromone induction for all experiments. If the p AGA1 -dPSTR Y is not induced properly, the experiment will be rejected. C. Correlation of the normalized expression level at 0, 20, 40, 60 min after the stimulus between the p AGA1 -dPSTR Y (y-axis) and p AGA1 (left), p SYN 3TT (middle) and the p FIG1 (right)-dPSTR R (x-axis). The Spearman correlation coefficient for each distribution is indicated in the lower right corner. D. Description of the metrics measured from a single cell trace of nuclear enrichment. The mean of the nuclear enrichment of the first 3 time points is used to quantify the basal level of expression of the trace. The difference between the maximum of the trace and the basal level represents the expression output (EO). When the trace overcomes the threshold set by the 20% of this EO added to the basal level, the response time (RT) is defined. E. To characterize individual single cell traces as not responding, weakly or strongly responding, the mean EO of all the cells of a reference strain is used. In the present case, the reference strain is the p AGA1 -dPSTR R construct which is used as a reference. Two criteria are used to define expressing cells. First, the last 5 points of the trace have to be significantly higher than the basal level (sign-test, blue, red, yellow traces). Second, the Expression Ouput of the trace has to overcome the expression threshold set at 20% ... |