Specificity of Qdot-RPPA. A. The regulation of AKT signaling pathway. Calyculin A is a phosphotase inhibitor leading to phospho-AKT accumulation. LY294002 inhibits PI3K phosphorylation therefore blocks the activation of downstream AKT signaling pathway. B. Cells were treated with inhibitors or serum as indicated to alter AKT activity. RPPA probed with either total AKT or phospho-AKT antibodies. All the samples were spotted in triplicates with 5 serial dilutions. C. Corresponding Western blots using the same lysates in B.