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University of Pittsburgh, Pittsburgh, PA, 15261

1 ARTICLES PUBLISHED IN JoVE

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Biology

The Ago2-miRNA-co-IP Assay to Study TGF- β1 Mediated Recruitment of miRNA to the RISC in CFBE Cells
Nilay Mitash 1,2, Joshua E. Donovan 1,3, Agnieszka Swiatecka-Urban 1,3
1Department of Pediatrics, UPMC Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, 2Department of Medicine, Division of Pulmonary, Allergy and Critical Care Medicine, 200 Lothrop Street, University of Pittsburgh, Pittsburgh, PA, 15261, 3Department of Pediatric Nephrology, University of Virginia, Charlottesville, VA, 22908

Here, we describe the Ago2-miRNA-co-IP assay designed to quantify an active pool of specific miRNAs induced by TGF-β1 in human bronchial epithelial CFBE41o- cells. This assay provides functional information on the recruitment of miRNA to the RNA induced silencing complex, quantified by qRT-PCR using specific miRNA primers and TaqMan hydrolysis probes.

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