Fig. 5.

Anti-miR-668 dampened EPO effect on AQP4 expression. (A) Astrocytes were transfected with antimiR-668 or anti-miRNA negative control. After 48 h of transfection, cells were treated with EPO (20 IU/ml) for 0, 2, 4, 8, and 24 h. Total RNA was extracted, and the miR-668 levels were determined using RT-PCR. The relative expression levels of miR-668 were determined with respect to the negative controls. The data shown here are from a representative of three experiments ± SEM. *p < 0.01, #p < 0.05 vs untreated group. (B) The corresponding AQP4 levels were also determined. Astrocytes were transfected with anti-miR-668 or anti-miRNA negative control. After 48 h of transfection, cells were treated with EPO (20 IU/ml) for 0, 24, and 48 h. Total protein was extracted and subjected to Western blotting to determine the AQP4 protein levels. The same blot was reprobed with anti-actin antibody for endogenous control. The data shown here are from a representative of three experiments