|- Cells were treated with Superoxide Detection Solution and then incubated for 30 min
To determine the level of superoxide production in cells expressing GFP-tagged GSK-3β, a Total ROS/Superoxide Detection kit (Enzo Life Sciences) was used according to the manufacturer's instructions. Cells were loaded with Superoxide Detection Solution and then incubated for 30 min. After washing the cells twice with wash buffer, the cells were observed by fluorescence microscopy. Fluorescence images of orange fluorescence, indicating superoxide, in GFP-positive cells at a magnification of 400× were analyzed by counting positive pixels per cell by image analysis software (Photoshop CS6, Adobe). Full paper
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