Fig. 5.

Detection of the DUX4 mRNAs in FSHD primary myoblasts. (a) Schematic representation of the DUX4 RNAs with alternative 3′ ends and the primers used for RT (no. 407) and PCR (nos. 222 and 407). (b) For controls, RT-PCR was performed on 3–4 μg of total RNA extracted from C2C12 cells transfected with the pGEM7Z vector containing either no insert (lanes 2 and 3) or the 13.5-kb genomic fragment of a patient with 2 D4Z4 units (pGEM42, lanes 4 and 5). RT-PCR was similarly performed on total RNA of control (N036, 9719, and C20) and FSHD primary myoblasts (F22, 5 D4Z4 units; M038, 7 units) either in proliferation (lanes 6–9) or in differentiation (diff) (lanes 10–13). RNA samples were incubated (+) or not (−) with DNase I and RT, as indicated. As a positive control (lane 14), PCR was performed on the pGEM42 vector present in a control sample (as in lanes 4 and 5) not treated with DNase I.