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. 2016 Nov 22;17(9):2354–2366. doi: 10.1016/j.celrep.2016.10.082

Figure 3.

Figure 3

Sox6 Is Required for Normal Nfix Function in Fetal Muscle

(A–D) Immunofluorescence with anti-Nfix (green) and anti-MyHC-I (red) antibodies on E16.5 muscle sections from WT (A and B) or Sox6-null (C and D) mice. Arrows indicate Nfix-positive nuclei in secondary (MyHC-I negative) fibers. Arrowheads indicate Nfix-positive nuclei in MyHC-I-positive fibers. Nuclei are counterstained with Hoechst. Scale bars, 25 μm. Higher magnifications of Nfix-positive fibers are shown in the insets.

(E) ChIP assay with anti-Nfix antibody on WT and Sox6-null E16.5 myotubes on negative control region (intergenic) located 15 kb upstream of MyHC-I TSS, the MyHC-I proximal promoter (−375 bp; MyHC-I promoter), and the Nfatc4 promoter region (−1.2 kb upstream of the Nfatc4 TSS) (∗p < 0.05; ∗∗p < 0.01; n = 2).

(F) Luciferase report assay on WT and Nfix-null differentiated fetal myoblasts (E16.5) transfected with control pGL3-basic, MyHC-I 408, and MyHC-I 408 m vectors (∗∗p < 0.01; n = 2).