Fig. 5. Effect of miR-29a on in vitro ischemia in astrocytes.

A, B. Dose-response of miR-29a levels to transfection with increasing amounts of miR-29a mimic (A) or inhibitor (B) in primary cultures of astrocytes, relative to control (Ctrl=1). Arrows indicate the dose we used for the following experiments. C. Representative immunoblots show PUMA protein levels in primary astrocytes without stress or after 6 h GD after transfection with control, mimic, or inhibitor. D. The graph shows the quantification of the Western blots. E. Effect of miR-29a mimic or inhibitor on astrocyte injury induced by 24 h glucose deprivation (GD). F. Effect of miR-29a mimic or inhibitor on cell injury induced by 7.5 h oxygen glucose deprivation (OGD) in primary astrocyte cultures. G. Effect of miR-29a mimic and inhibitor on mitochondrial membrane depolarization in astrocytes subjected to 3 h GD. Depolarization is indicated by decreased TMRE fluorescence. H. Effect of miR-29a mimic or inhibitor on ROS in astrocytes subjected to 3 h GD. Increasing HEt fluorescence indicates increasing ROS. Fluorescence values are normalized to the starting fluorescence = 1.0. Ctrl: transfection control. All experiments were performed 3 times in triplicate. *P<0.01 compared to Ctrl.