mRNA expression profiling after Ago2-immunoprecipitation (IP) in unstimulated and TGF-β1-stimulated primary parenchymal lung fibroblasts of two control subjects

To identify the target gene repertoire of miRNAs (i.e. the miRNA-targetome) of unstimulated and TGF-β1-stimulated primary parenchymal lung fibroblasts, Ago2-IP was performed followed by mRNA expression profiling. The Ago2-IP was performed on the unstimulated and TGF-β1-stimulated primary parenchymal lung fibroblasts of two control subjects. The RNA of the Total (T) fraction and the immunoprecipitated (IP) fraction were isolated using miRNeasy Mini Kit and miRNeasy Micro Kit, respectively. Subsequently, the RNA was labeled with Cyanine 3 (Cy3) and Cyanine 5 (Cy5) in a dye-swap experiment and cRNA was generated which was purified using the RNeasy Mini Kit. The labeled and amplified cRNA was then used to perform the mRNA expression profiling using the G3 Human Gene Expression 8x60K v3 Microarray.