Figure 2.
GSH measurements in uninfected neutrophils cultured in the presence and absence of NAC or L-GSH (n = 5). GSH levels were measured in isolated neutrophils from healthy subjects that were treated with GSH-enhancing agents such as NAC and L-GSH, by spectrophotometry using an assay kit. Briefly, neutrophils (3 × 105) were lysed at 24 hours after treatment with ice cold 5% 5-sulfosalicylic acid dehydrate solution. Supernatants collected after centrifugation were analyzed for total GSH (a) and GSSG (c) as per manufacturer's instructions. rGSH was calculated by subtracting measured GSSG concentrations from the measured total GSH concentrations (b). All GSH measurements were normalized with total protein levels. Proteins in the cell lysates of neutrophils were measured by Bradford's method using a Coomassie protein assay reagent. (d) illustrates ratio of rGSH to GSSG expressed as percentage of total GSH in neutrophil lysates. Graph shows mean ± standard error.