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Fig. 4 | Skeletal Muscle

Fig. 4

From: Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice

Fig. 4

In non-muscular tissues, enhanced DUX4 expression in D4Z4-2.5/Dnmt3bMommeD14 mice is limited to the secondary lymphoid organs. a DUX4 transcript levels were enhanced as measured by RT-qPCR in the inguinal lymph nodes and the spleen of D4Z4-2.5/Dnmt3bMommeD14 mice (postnatal day 15) while the other tested tissues remained unaffected. Each dot represents one mouse, and the error bars denote the SEM of the biological replicates. Statistical analysis was performed with a Student’s t test (brain, heart, lymph nodes, muzzle skin, spleen, testis) or a Mann–Whitney U test (bone marrow, thymus). *P < 0.05; ***P < 0.001. n.s. = not significant. 2.5 = D4Z4-2.5; MD14 = Dnmt3bMommeD14. b Transcript levels of the target gene Wfdc3 were upregulated in the inguinal lymph nodes and the spleen of D4Z4-2.5/Dnmt3bMommeD14 mice at postnatal day 15 as measured by RT-qPCR. Each dot represents one mouse, and the error bars denote the SEM of the biological replicates. Statistical analysis was performed using a Student’s t test (lymph nodes) or Mann–Whitney U test (spleen). **P < 0.01. 2.5 = D4Z4-2.5; MD14 = Dnmt3bMommeD14. c With RT-qPCR we showed that Cxcl12 and Podoplanin transcripts (stromal cell markers) were highly enriched in the stromal cell fraction while CD19 transcripts (B cell marker) were enriched in the immune cell fraction, confirming a successful separation of both fractions. DUX4 transcripts were highly enriched in the stromal cells, while the immune cells showed a low expression. In both fractions, DUX4 transcript levels were elevated by the Dnmt3bMommeD14 variant (not significant). Statistical analysis was performed by one-way ANOVA. Each bar represents at least four biological replicates, and the error bars denote the SEM. **P < 0.01; ***P < 0.001; ****P < 0.0001. n.s. = not significant. 2.5 = D4Z4-2.5; MD14 = Dnmt3bMommeD14

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