Fig. 3

Antisense sODN binds the target sequence. Panel a Relative location of four primers on HDAC5 mRNA. Both miD2861 and hdac5AS2 are in AS orientation. Panel b Modified gel shift assay. We mixed Rhd-miD2861 with one sODN of four different primers (lane 1: the passenger of miR-2861, lane 2: matured miR-2861, lane 3: Rhd-miD2861, and lane 4: cDNA of HDAC5 mRNA at the miR-2861 binding site [sODNhdac5]). Binding of miD2861 slowed mobility during gel electrophoresis. Panel c Differential retention assay. We transfected Cy2-AS first for 3 h, washed, then transfected Rhd-sODN. Eighteen minutes later we washed and changed medium again, then took photography immediately and at 2 h later. Sense sODN was excluded preferentially at 2 h