We utilize an immortal endothelial cell line and various agonists to observe their effects on Src phosphorylation. However, our protocols seem to induce high basal levels of Src phosphorylation. Given that these cells are a hybridoma of HUVEC and cancer cells, they may naturally exhibit elevated levels of Src and its activated form. I have heard that the process of removing the starvation medium (which contains 0.5% FBS) and adding the incubation medium (also with 0.5% FBS) can stimulate Src phosphorylation. Is this true? If so, are there any studies that discuss this? I have been unable to find any relevant literature.