RIP assay was performed as described previously [24 (link)]. Briefly, PANC-1 cells were transfected with LINC00346 and miR-188-3p and resuspended in lysis buffer. Cellular lysates were incubated with Protein G sepharose beads conjugated with anti-Ago2 (Abcam) or anti-IgG (Abcam) for 4 h at 4 °C. The immunoprecipitates were treated with DNAse I and proteinase K for 20 min at room temperature. Co-precipitated RNA was recovered and subjected to qRT-PCR analysis.
Free full text: Click here
