Figure 2. Validation of miRNA expression changes and their dependence on glutamine synthesis and NADPH-oxidase activation in ammonia-exposed astrocytes.

(A/B) Cultured rat astrocytes were exposed to NH₄Cl (5 mmol/l) or were left untreated (control) for 48 h before RNA was isolated and analysed for miRNA expression changes by Agilent miRNA Array (A) or miQPCR (A/B) as described in materials and methods. (A) Validation of miRNA expression changes by miQPCR. (B) Glutamine synthetase and NADPH-oxidase dependence of miRNA expression changes in NH₄Cl (5 mmol/l, 48 h)-exposed astrocytes. Where indicated astrocytes were treated with MSO (3 mmol/l) or apocynin (300 μmol/l). *statistically significantly different to the respective control (untreated or inhibitor-treated). n.s.: not significantly different to the respective control (untreated or inhibitor-treated). Data are represented as log₂ ratios from 3–10 independent experiments.