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. 2006 Jul 19;34(12):3421–3433. doi: 10.1093/nar/gkl390

Figure 6.

Figure 6

Pactamycin elicits latent splicing in wild-type CAD RNAs. (a) Cells were transfected with CAD mini gene constructs as indicated. The presence of wild-type or mutated start and stop codons in each construct is marked by + or −, respectively (see Supplementary Table 1S). Twenty-four hours post-transfection, cells transfected with CAD1 (lanes 3–6) were treated with pactamycin (lanes 3–5) or CHX (lane 6), as indicated. RT–PCR products were analyzed on a denaturing 10% polyacrylamide gel. (b) Quantitative analysis of latent splicing was carried out as described in Figure 5 using RNA from cells transfected with wild-type CAD1 and CAD2 and treated with 3 μg/ml pactamycin for 4 h. Error bars represent SE of three independent experiments. (c) Transfection experiments were performed with CAD1 for 24 h. The cells were treated for 2 h with pactamycin (pact; 3 μg/ml; lane 3), tetracycline (tet; 20 μg/ml; lane 4) and hygromycin B (hyg B; 20 μg/ml; lane 5) and RNA was extracted. RT–PCR products were analyzed on a denaturing 5% polyacrylamide gel.