Identification of host miRNAs that may limit human rhinovirus replication

Figure 5 Antibody titration for AGO2 co-IP. These experiments were performed in order to determine the most convenient antibody concentration to use for (A) THP-1 (used as additional control) and (B) BEAS-2B cells: 1μg was chosen as it would give the highest enrichment of both microRNAs and target mRNAs. These results show that microRNAs are very tightly associated with AGO2 protein, and confirm that microRNA-regulated mRNAs are also considerably co-purified. Previous work from our group has shown that PU.1, SMAD2 and IL13RA1 are targeted by miR-155[45,62,89] while COL1A1 has been shown to be targeted by miR-29b elsewhere[90]. IgG refers to the negative control antibody, AGO2 refers to the anti-AGO2 antibody used. Plotted values represent the average of qPCR duplicates from one experiment.