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. 2021 Oct 4;10:e65672. doi: 10.7554/eLife.65672

Figure 7. Myogenin preferentially induces microtubule-organizing center (MTOC)-associated isoforms of Syne1 and Akap6.

(A, C) Schematic representation of the murine Syne1 (A) and Akap6 (C) gene and derived transcripts. Exons are indicated by gray rectangles and the first exon of each transcript is marked by color. E-boxes (myogenin consensus sites) inside putative promoters are indicated as yellow boxes and small black arrows mark the primers used for qPCR. (B, D) Myogenin chromatin immunoprecipitation (ChIP) from doxycycline (Dox)-stimulated MYOG-mScarlet cells followed by qPCR for the indicated E-boxes shows that myogenin preferentially binds the promoter regions upstream of Syne1 α-isoform and Akap6 β-isoform transcripts. (E, F) Luciferase assay testing the activity of the indicated Akap6 (E) or Syne1 (F) promoters in the presence of GFP or myogenin-GFP (MYOG-GFP). (G) Overexpression of nesprin-1α-mCherry alone or together with AKAP6β-GFP in undifferentiated (myogenin-negative) C2C12 myoblasts. Co-expression of nesprin-1α and AKAP6β is sufficient for nuclear envelope recruitment of endogenous PCM-1. Scale bars: 20 µm. Data (B, D–F) are represented as individual biological replicates (n = 3), together with mean ± SD. ns: p>0.05; **p<0.01; ***p<0.001.

Figure 7—source data 1. Underlying data for graphs in Figure 7B, D–F.

Figure 7.

Figure 7—figure supplement 1. Ectopic co-expression of nesprin-1α and AKAP6β is not sufficient for microtubule-organizing center (MTOC) function at the nuclear envelope.

Figure 7—figure supplement 1.

Proliferating (myogenin-negative) C2C12 myoblasts were transfected with nesprin-1α-mCherry and AKAP6β-GFP. Two days after transfection, microtubule regrowth assays were performed. Scale bar: 10 µm.