Figure 3

Northern blot analysis of the host gene encoding miR-143 and - 145. Total RNAs (5 μg for human normal tissues, cancer tissue, and cancer cell lines [human stomach tumor RNA was purchased from Takara]) were separated on a 1%(w/v) formaldehyde agarose gel and then blotted onto Hybond N+ nylon membranes (Amersham Biosciences, Piscataway, USA). RNA probes were synthesized from template cDNAs by using a MAXIscript kit (Ambion, Austin, USA) incorporating Digoxigenin-11-UTP (Roche, Penzberg, Germany). Northern blots were hybridized with the AKF1-10 clone (A, upper panel; refer to Fig. 2), 143-145 clone (A, 2nd panel), GAPDH (A, 3rd panel; B, middle panel), and 28 S rRNA (A and B, under panel). The primary transcript is marked by the open arrow and other transcripts are marked by closed arrows.