Figure 4.

Cryptic splicing does not occur in the dicistronic transcript containing ACLY 5′ UTR. (A) Dicistronic constructs used to transfect HepG2 cells are depicted. The arrows indicate the positions of the primers CSFor and CSRev used for RT-PCR and reported in Table 1. RT-PCR analysis was performed as additional test to rule out cryptic splicing within the ACLY 5′ UTR in the intercistronic region. (B) RNA was extracted by HepG2 cells transfected with the dicistronic constructs and used as a template for RT-PCR. The amplimers containing the RL and the intercistronic region were analyzed on ethidium-bromide-stained agarose gel. The result is from a representative experiment (n = 3).