CYTO-ID® Autophagy detection kit

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	Mouse pancreatic NIT cells when treated with Atorvastatin or Pravastatin show increase in

Protocol tips
Mouse pancreatic NIT cells when treated with Atorvastatin or Pravastatin show increase in

Publication protocol

The 2.5 × 105 cells were seeded in a 3.5 cm confocal dish and pretreated with various inhibitors before statin treatment. Following a 48 h treatment with statin, the cells were washed with PBS to remove dead cells and serum proteins. Immediately, treated or untreated NIT-1 cells were stained with an antibody against rabbit monoclonal AIF (1 : 400, Cell Signaling) or CYTO-ID Autophagy Detection Kit (Enzo Life Sciences, Farmingdale, New York, USA) and imaged via confocal microscopy using an Olympus FV1200 microscope (Tokyo, Japan). The images were converted to 8-bit grayscale and analyzed by the ImageJ software. The integrated density (IntDen) provided a measure of intensity proportional to total volume and was calculated using area. Values from six random images for each group were averaged for comparison.

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