Ago2-RIP-chip in Dicer WT and KO macrophages

To determine the spectrum of miRNA targets regulated following Dicer deletion, we performed argonaute 2 (AGO2)-RNA Immunoprecipitation (RIP)-microarray in bone marrow-derived macrophages (BMDMs) from LysM-Cre/Dicerflox/flox/Apoe–/– and LysM-Cre/Dicerwt/wt/Apoe–/– mice. This analysis combined with miRNA profiling in Dicer wild type (WT) and knockout (KO) BMDMs may help to identify the miRNA targets regulated by Dicer deletion. Bone marrow cells were harvested from femora of 6-8 week old male LysM-Cre/Dicerflox/flox/Apoe–/– or LysM-Cre/Dicerwt/wt/Apoe–/– mice, re-suspended in DMEM-F12/10% FCS/10% L929-conditioned medium, and cultured for 7 days to differentiate into primary macrophages. RNA was isolated before and after AGO2-RIP or IgG control-RIP using Trizol.