Parallel profilling of signaling pathways

A method for identifying signaling pathways that regulate the expression of a test gene, the method comprising

• (a) combining each of n cell cultures with each of n compounds, where a different compound is combined with each cell culture,
• wherein the n cell cultures are capable of expressing the test gene, and wherein signaling pathways affected by each of the compounds are known;
• (b) measuring expression of the test gene in each of the cell cultures to identify compounds (“active compounds”) that affect expression of the test gene in the cell culture;
• (c) combining cell cultures with an active compound identified in step (b) and to at least a portion of the n compounds, where a different compound is combined with each cell culture;
• (d) measuring expression of the test gene in each of the cell cultures of step (c) to identify compounds that negate the effect that the active compound has on expression of the test gene (“negating compounds”);
• and
• (e) identifying the signaling pathways that are affected by a negating compound (“test gene pathways”), wherein the test gene pathways are signaling pathways that regulate the expression of the test gene.