Skip to main content
. 2008 Dec 18;3(12):e3968. doi: 10.1371/journal.pone.0003968

Figure 7. Expression of LacZ-(CUG)400 RNAs is insufficient to dysregulate IR and cTNT splicing in human myoblasts.

Figure 7

Panel A: DMPK 11-15(CTG)5 or 300 (a), GFP-DMPK 3′UTR (CTG)5 or 400 (b) and LacZ-(CTG)0 or 400 (c) cassettes under the transcriptional control of the cytomegalovirus (CMV) promoter are shown. Panel B: Nuclear DAPI staining of human normal myoblasts expressing DMPK11-15(CTG)5 (a), DMPK 11-15(CTG)300 (b), GFP-DMPK 3′UTR(CTG)5 (c), GFP-DMPK 3′UTR(CTG)400 (d), LacZ-(CTG)0 (e), LacZ-(CTG)400 (f) cassettes are shown. The mutant transcripts encoding the expanded CUG tracts were detected by hybridization with a (CAG)10-Cy3 probe. CUG RNA foci are observed primarily within the nucleus in normal myoblasts expressing DMPK11-15(CTG)300 (red signal; b) and GFP-DMPK 3′UTR (CTG)400 (red signal; d). CUG RNA foci are observed in the cytoplasm (red signal; f) in normal myoblasts expressing the LacZ-(CTG)400 cassette. Normal myoblasts expressing DMPK11-15(CTG)5 (a), GFP-DMPK 3′UTR(CTG)5 (c), and LacZ-(CTG)0 (e) constructs did not show RNA foci. Panel C: IR and cTNT RNA splicing in myoblasts expressing the indicated cassettes are shown. Synthesized cDNAs (150 ng) were subjected to RT-PCR analysis using the IR and cTNT primers described in Methods. GAPDH RNA was amplified in parallel as an internal control. The experiments were carried out in triplicate. Representative panels are shown in Panel C and the results are tabulated in Table 4.