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Figure S3. Cleavage sites on B-ind1 or Pola1 mRNA. (A) The sequence of B-ind1 mRNA (available from GenBank/EMBL/DDBJ under accession no. NM_021345) is shown with the cleavage sites marked. Downward-facing closed arrowheads are obtained from miR-206 transfected cells and upward-facing, open arrowheads are from GL2 control transfected cells. Asterisks and black ovals are from miR-1 and -133 transfection, respectively. Number of arrowheads indicates frequency of cloning the cleavage product. Although miR-133 induced far fewer cleavage sites (Fig. 7, E and F), a few were cloned and sequenced to demonstrate that the distribution of cleavage sites is different from that obtained by miR-1 or -206. Underlined are the predicted target sequences by miRanda program, with -10 kcal/mol as ΔG threshold. The two target sites mutated in Fig. 7, B and C (M1 and M2) are in bold. The primer sequences used for the RACE-PCR are indicated by arrows. Solid arrows indicate primers for cDNA synthesis (2683-2655 and 1500-1479 for P1 and P2 set in Fig. 7 A, respectively), dashed arrows indicate primers for the first-round PCR (2658-2633 and 1481-1459), and dotted arrows indicate primers for the second-round PCR (2637-2611 and 1454-1430).