(A) RNA-seq read coverage of the SRSF3 gene in control and DUX4-expressing myoblasts. 54-1, immortalized myoblasts; MB135, primary myoblasts. Purple shading, NMD-inducing cassette exon. Red stop …
(A) Poly-caspase activity (red) following transfection with a control siRNA or siRNA against TP53 40 hr after lentiviral infection. Box plot, percentage of nuclei with poly-caspase granules …
(A) Relative mRNA levels of NMD factors in DUX4-expressing vs control myoblasts (54-1 cells). Red, up-regulation by ≥1.5-fold. (B) Immunoblot for NMD factors UPF1, SMG7, and UPF3B in DUX4-expressing …
(A) Schematic of the DUX4 mRNA. Intron 2, constitutively spliced intron within the 3′ UTR. Black, coding sequence; purple, 5′ and 3′ untranslated regions (UTRs). Red stop sign, termination codon. (B)…
(A) Schematic of potential double-negative feedback loop between DUX4 and NMD, in which DUX4 inhibits NMD and NMD degrades DUX4 mRNA. (B) Levels of endogenously transcribed DUX4 mRNA following …
Primer sequences used for cloning and real-time PCR assays. List of primers used for cloning and real-time PCR. GENE ID, Ensembl ID for the tested genes. RPL27 and SRP14: housekeeping genes. HNRNPD, SRSF2, and SRSF3: inclusion primer sets measure levels of endogenously produced NMD-degraded isoforms. Endogenous DUX4 mRNA was measured with primers specific to its 3′ UTR (RKB_929 and RKB_930). Expression from chimeric Gl-DUX4-Intron2(+) and Gl-DUX4-Intron2(−) constructs was measured with primers specific to the spliced β-globin region.