Figure 6

Effect of PNA2406 on the splicing of introns 2, 3 and 4. JAR cells were transfected with PNA2406 at 2 μM for 24 h and subjected to RT-PCR analysis using the extracted total RNAs. N: no PNA treatment. P: treated with PNA2406 (2 μM). (A) Location of the PNA2406 and splice sites analyzed ((a), (b), (c), (d) and (e)) are indicated. (B) The structures or names of RT-PCR products are shown on the right of each picture. Primer sets for the detection of each splice sites (in Figure 6A) by RT-PCR (product length) are as follows: (a), A (161 bp) and H (β-actin, 223 bp); (b), D (169 bp) and H (β-actin, 223 bp); (c), E (251 bp) and I (β-actin, 166 bp); (d), F (151 bp) and H (β-actin, 223 bp); (e), G (215 bp) and I (β-actin, 166 bp). The numbers under the figure indicate the relative amount (normalized to β-actin) of the target mdm2 splicing variants. (C). Primers, Exo2S and Exo4AS in Table 2, were used for RT-PCR for the detection of exon3-skipped form. Locations of the splice inhibited form (inhibited (318 bp)), normal splice form (normal (197 bp)) and exon3-skipped form (skipped (112 bp)) although the last one (skipped form) was not visible.