Anti-Argonaute RIP-Chip shows that miRNA transfections alter global patterns of mRNA recruitment to microribonucleoprotein complexes

(A) Clustering analysis of RIP-Chip microarray expression data indicating that upstream miRNA transfections cause consistent changes in the mRNAs recruited into the miRNP. Anti-AGO co-IPs of H4 cells were processed for Affymetrix Gene 1.0 ST microarrays. This heat-map representation of the microarray expression profiling after transfection with various miRNAs and RIP-Chip comprises 15 different biological replicates. The abscissa refers to the same 15 biological replicates in the same order. The analysis indicates that the transfections produced consistent results with between-group variability exceeding within-group variability. (B) Endogenous expression levels of selected miRNAs in H4 cells. Levels of selected brain enriched miRNAs were evaluated as described previously (Wang et al. 2008b) by miRNA microarray analysis of H4 cell RNAs.