(A) DeMAND requires both a regulatory network and a set of gene expression profiles from compound perturbed and control samples, as an input.
(B) DeMAND evaluates the dysregulation of each interaction in the regulatory network.
(C) To evaluate interaction dysregulation co-expression scatter plots for the two interacting genes are smoothed using a Gaussian Kernel method to generate an interaction probability density. The probability density difference before and after compound perturbation is evaluated using the KL-divergence. The top example illustrates no change in probability density (i.e., no dysregulation). The other three examples illustrate various examples of compound dysregulation, including correlation inversion, gain, and loss (top to bottom, respectively).
(D) The statistical significance of the KL-divergence is assessed by gene pair shuffling.
(E) The global dysregulation of each gene is determined by integrating the p-values of all its network interactions, while accounting for their dependencies (see also Figure S1 and Extended Experimental Procedures).
(F) DeMAND produces a list of all network genes and the statistical significance of their dysregulation.