miR-96 binds to the second predicted binding site (96.2) in the 3'UTR of FOXO1 to decrease the protein level.

<p><b>A</b>. There are two predicted miR-96 binding sites in the FOXO1 3'UTR sequence. The seed region of the mature miR-96 and the predicted binding sites in the 3'UTR sequence are underlined and bold. Locations of the binding sites are according to Targetscan, (Release 6.2, June 2012). <b>B</b>. 22Rv1 cells were co-transfected in triplicates with miR-96 mimic and a target site blocker for binding site 96.1 in three concentrations. FOXO1 protein level did not increase when binding site 96.1 was blocked. <b>C</b>. Blocking binding site 96.2 with 6x the concentration of target site blocker compared to the miR-96 mimic resulted in a significant increase of the FOXO1 protein level (p=0.0118). FOXO1 protein levels were normalized to GAPDH. Error bars show standard error of mean.