The first experiment that they did, the experiment involving adding hemoglobin to lung epithelial cell culture answer the question by modeling what happens in the body If there were Hemoglobin S, Hemoglobin SE, and hemoglobin A, normal hemoglobin) was floating in the bloodstream. In the experiment, they measured the permeability by having the epithelial cells grow on a membrane rather than in a flask. Then they measured the permeability of the culture by adding media containing dextran on the top layer and measuring how much dextran there were in the bottom layer. At this stage, they also added hemoglobin to the top layer to see if the hemoglobin had any effect on the permeability of the cell layer. The cells were then washed and immunoblotting and microscopy were done on the cells to visualize how the hemoglobin had an effect on the cells. The researchers also measured the amount of oxidative stress that the cells had gone through by measuring the epithelial lipid hydroperoxide levels. Mitochondrial Bioenergetic and glycolytic levels were also measured to assess the damage in the cells. The procedure of this experiment is simple in the sense that they had a model and exposed the model to the same environment as those that would be found in sickle cell disease patients, and assessing if there were any damage that the cells sustained. While this experiment does not exactly measure the dysfunction that is present in the hemoglobin (as for example, exactly how and why it is dysfunctional), it measures what the hemoglobin dysfunction causes in other cells (what does the dysfunction of the protein does to other cells?).
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