FIGURE 4.
Functional effects of mirtron-mediated DMPK knockdown in DM500 cells. DM500 cells were transfected with 1 μg of NAD, DMPK Mirt 5, or DMPK Mirt 5US and differentiated 2 d after transfection. After 7 d in differentiation medium, RNA was harvested. (A) Representative GFP fluorescence and merged brightfield and eGFP images of differentiated DM500 before harvest. (B) qPCR of DMPK cDNA normalized to mGAPDH cDNA. (C) Representative RT-PCRs of Serca-1 using primers flanking exon 22. Sequestration of MBNL1 leads to exclusion of exon 22. The graph below shows the relative ratio of cDNA with exon 22 to cDNA without exon 22. (*) p < 0.05 versus NAD control.
