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. 2017 Mar 17;36(9):1199–1214. doi: 10.15252/embj.201695988

Figure 3. Molecular consequences of loss of miR‐22 in muscle tissue.

Figure 3

  • A, B
    The direct miR‐22 target protein ERα is upregulated in muscle tissue of miR‐22 (KO) male mice, but not in fat tissue (muscle: WT, n = 5; miR‐22 KO, n = 5; epididymal fat: WT, n = 4; miR‐22 KO, n = 4).
  • C
    ERα is not significantly regulated in muscle tissue of female mice (WT, n = 5; miR‐22 KO, n = 5). Original Western blot data are included in Appendix Fig S1. ERα/GAPDH ratios were normalized to the mean of WT values.
  • D, E
    Luciferase reporter assays in C2C12 proliferating myoblasts or differentiating myocytes demonstrate that the WT miR‐22 target site originating from the 3′ UTRs of ERα mRNA represses the activity of luciferase upon transfection of pre‐miR‐22 (miR‐22), but not upon transfection of scrambled control miRNA (scr) and this is not observed with a mutant version (mut) of the target sites. Myocytes express miR‐22 endogenously, and this results in additional repression of luciferase under control of WT target site compared to mutant target site in myocytes (n = 3 independent transfections each).
Data information: Data are presented as mean ± SEM. **P < 0.01, two‐tailed unpaired t‐test.