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8) after multiple testing correction (Bonferroni) are considered buffering. (1) single criterion for ��significant change in mRNA expression�� in a mutant is applied EAI045 clinical trial throughout the entire study: the average change in expression of a gene must be more than 1.7-fold with a p value of less than 0.05 versus the collection of WT cultures processed in parallel. Complete redundancy: No significant changes (i.e., less than 8 genes with p? 1.7) in both singles and a significant change in the double (8 or more genes). Quantitative redundancy: When thresholded for p? 1.7 there is a more than 95% overlap in the genes reported as changed in the double versus the two singles combined but that is accompanied by a significant shift in the magnitude of fold changes (Mann-Whitney test, p?GUCY1B3 8 or more genes change significantly in the double versus either of the two singles in a manner other than complete or quantitative redundancy. Known binding motifs for Ste12 (TGAAC) and Tec1 (CATTCYY) were obtained from literature (Madhani and Fink, 1997) in IUPAC-IUB notation (http://www.iupac.org/). The binding motifs were matched against the upstream sequence (?800bp to ?1bp upstream of transcription start site) of the genes significantly upregulated in the fus3�� and unchanged in the fus3�� kss1�� mutant or significantly downregulated in the fus3�� kss1�� mutant and unchanged in the fus3�� A-1331852 mutant using the dna-pattern tool on the Regulatory Sequence Analysis Tools (RSAT) website (http://rsat.ulb.ac.be/rsat/) ( van Helden et?al., 2000). Given 4 nodes and no self-edges between nodes, a total of 528,576 connected network topologies exist. For the sake of biological relevance as well as computational tractability we constrained the models to those having at least two edges from the regulator nodes (K1 and K2) to the responder nodes (R1 and R2). The number of incoming edges on any node was limited to a maximum of two, and their influence on the node could be a Boolean AND or Boolean OR (the NOT function is a property of an inhibitory incoming edge, not of the node). If one or more nodes of the graph have at most two incoming edges for a particular topology, all possible combinations of AND and OR-functions on each of these nodes were generated. The total number of models thus obtained was 794,176. Boolean network model simulations have been performed using the BooleanNet software application (http://booleannet.googlecode.com) (Albert et?al., 2008). Synchronous Boolean simulations were run for all possible initial states (True or False) of K2, R1 and R2 in 15 consecutive steps, K1 is set to True in all cases, resulting in 8 possible initial states of a particular Boolean network model. For all initial node states, the WT, k1��, k2�� and k1�� k2�� genotypes were tested (32 simulations per model) for compatibility with our experimental results.