Dataset used for parameter estimation.

Five different cell lines that have a perturbation in the level of a key signal transduction protein were constructed by shRNAi lentiviral infection. The calcium response from these cell lines in addition to the wild-type cell line were used to fit relevant parameters in the model. Because shRNAi does not entirely remove the protein product, the fraction knockdown was estimated by qRT-PCR and by Western blot analysis. The standard error (se) was computed for each estimate and the upper and lower confidence intervals were computed as ±3·se. The knockdown confidence intervals are used in the GPCR model to construct prediction confidence intervals for the calcium response. Where several cell lines were constructed for each knockdown, the best was selected and reported in parenthesis. The sample size for each knockdown-ligand dose combination is shown in the last 6 columns.