Mitochondrial fission and fusion were determined in VSMCs transduced with adenovirus expressing mtGFP (MOI of 50) for 48 hours. For automated morphometry, images were processed using NIH ImageJ software, with the plug-ins involving either “rolling ball” background subtraction or deblurring by two-dimensional deconvolution with a computed point spread function. Using a custom-written NIH ImageJ macro provided by S. Strack (University of Iowa), processed images were converted to binary (black and white) images by autothresholding, and mitochondrial particles were analyzed for length or form factor [perimeter 2/(4 × π × area)] (47, 48). The parameters for form factor are set with a minimum value of 1 for perfectly circular mitochondria.

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