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If I mess up the timing once, have I ruined my entire protocol

Regarding oxidative stress, the enzymatic systems that contribute the most to the generation of ROS include the proteins that are bound to the plasma membrane, such as the family of NADPH oxidases [17,18], the enzymatic systems that participate in the lipid metabolism within peroxisomes and the activity of various cytosolic enzymes such as cyclooxygenases

Figure 4 GSH-Mediated Modulation of Stress-Associated Hormones Maintains Cellular Hormone Balance in Cd-Stressed Oilseed Rape Plants The levels of the stress-inducible hormones abscisic acid, jasmonic acid, and salicylic acid were determined to assess the influence of glutathione on hormone homeostasis in cadmium-exposed plants