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. 2014 Jan 2;9(1):e84294. doi: 10.1371/journal.pone.0084294

Figure 2. Increased NHE1 activity in hippocampal astrocytes following OGD/REOX.

Figure 2

A. pHi changes were determined by BCECF-AM dye in astrocytes subjected to NH4 +/NH3 prepulse-mediated acid-loading. Either normoxic astrocytes (left panel) or astrocytes at 2 h OGD (right panel) were exposed to 30 mM NH4Cl3 (a–c), then returned to standard HCO3 –free HEPES-MEM solution (c–e). After an initial acidification, pHi recovery followed (d–e). The pH recovery rates were determined at ∼ 6.5 to normalize for the allosteric regulation of H+ on NHE1 activity. A slope of the pH i changes following the prepulse was calculated as pHi recovery rate (solid or dashed line). For HOE 642 treatment, the drug was present throughout the experiment. B. pHi (left panel) and pHi recovery rates (right panel) were summarized under normoxic control, 2 h OGD, or 1, 5, or 24 h REOX conditions. C. Summary data of NHE1-mediated recovery rates under normoxic and OGD/REOX conditions. The values of ∼20 cells from each coverslip/culture were averaged. The n values were the number of cultures under each condition and indicated as normoxia (9), normoxia+HOE 642 (6), 0 REOX (5), 0 REOX+HOE 642 (4), 1 h REOX (3), 1 h REOX+HOE 642 (4), 5 h REOX (5), 5 h REOX+HOE 642 (4), 24 h REOX (4) and 24 h REOX+HOE 642 (3). Data are expressed as mean ± SEM. *p<0.05 vs. corresponding untreated. # p<0.05 vs. corresponding untreated.