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Immunoblots were imaged on a BioRad Pharos Plus Molecular Imager using the Cy5 setting (635 nm laser excitation, 695 nm emission filter).
Supernatant and pellet fractions were separated on sodium dodecyl sulfate polyacrylamide (SDS-PA) gels, stained with Coomassie blue and scanned in a BioRad Molecular Imager using the QuantityOne software.
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The bands were visualized with a PhosphorImaging system (Bio-Rad, Molecular Imager FX) using the manufacturer's software Quantity One, version 4.3.0.
Radioactivity was imaged and quantified by phosphor imaging with a Molecular Imager FX (Bio-Rad Laccompanying, using accompanying Quantity One software.
Chemiluminescent signal was generated by adding 10 mL of SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific), and the blot was imaged using Molecular Imager Gel Doc XR+ System (Bio-Rad).
The next day, secondary antibody was applied and the signal was visualized on a Molecular Imager ChemidocXRS (Bio-Rad Laboratheies) using the Pierce Supersignal West Pico chemiluminescent substrate (Thermo Scientific).
The gels were scanned using BioRad Molecular Imager FX and image analysis performed using ImageJ software http://rsb.info.nih.gov/ij.
Gel images were acquired by using Molecular Imager ChemiDoc XRS (Biorad) and analyzed by using Kodak Molecular Imaging Software 4.0 or IMAGEJ 1.40 g (Wayne Rasband, NIH).
A Bio-Rad Molecular Imager FX was used to quantify individual bands.
Radioactive labeling was visualized using a Pharos Fx Plus Molecular Imager (BioRad) and quantitated using ImageLab software (BioRad).
Gels were imaged using a Molecular Imager Gel Dox XR System (Bio-Rad Laboratories, Inc).
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