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Fig. 8. JP-45 gene silencing in differentiated C2C12 myotubes. (A) Total RNA was extracted from transfected and differentiated C2C12 cells and converted into cDNA. The cDNA encoding JP-45 and ß-actin was amplified by PCR. Amplified DNA obtained from 50 ng or 100 ng RNA was separated on a 7.5% acrylamide gel (JP-45, top panel) or a 1% agarose gel (ß-actin, bottom panel). (B) Microsomal proteins from transfected and differentaiated C2C12 cells were prepared, separated on a 10% SDS-PAGE, blotted onto nitrocellulose and probed with anti-JP-45 Abs (central panel) or commercial anti-ß-actin Abs, followed by peroxidase-labelled secondary Abs. Immunoreactive bands were visualized by chemiluminescence. Panel on the right shows blotted proteins stained with Ponceau Red. Results are representative of experiments carried in three different transfection experiments.
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