hi
i intend to study the phosphoproteome changes upon exposure of a glomerulus cell to increase glucose concentration. diabetic conditions have been shown to induce apoptosis in these cells in previous literature search but at the same time i am interested in comparing the phosphoproteome of cells exposed to glucose to maybe study the signalling changes in diabetic conditions.
how do i proceed if it causes apoptosis?
Apoptosis does not happen instantaneously. You may have to empirically select a few time points, post-hyperglycemic exposure, and compare the patterns of phophorylated proteins on 2-D gels.
If you find obvious differences, proceed further to identify the target proteins.
If you will properly select a concentration of glucose, you will avoid the risk of apoptosis. We worked with 30 mM of glucose using various cell types and there was no significant apoptosis. However, be careful because glucose may induce at this concentration accelerated cell senescence which will affect the phosphoproteome.
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