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. 2018 Mar 26;37(8):e98239. doi: 10.15252/embj.201798239

Figure 4. mTORC1 is a key downstream mediator of p110α in regulating quiescence exit in adult MuSCs.

Figure 4

  1. Age‐matched adult Ctrl, iKO, and idKO mice (n = 3 in each group) were first treated with Tmx as described in Fig 1. MuSCs were isolated by FACS and cultured with EdU for 36 h. Cells were fixed and subjected to staining for EdU (green) and p‐S6 (red). The nuclei were counterstained by DAPI (blue).
  2. Quantification of EdU+ or p‐S6+ MuSCs from (A) by counting ˜600 MuSCs/mouse.
  3. Representative histograms (top) showing the mean FSC‐A values and dot plots (bottom) showing the percentage of Vcam1+ MuSCs from the Ctrl, iKO, and idKO mice.
  4. FACS‐isolated MuSCs from the Ctrl, iKO, and idKO mice (without Tmx) were cultured for 2 days followed by 2 days of 4‐OHT treatment. Whole‐cell lysates were subjected to Western blotting.
  5. Freshly isolated single myofibers (n > 20) from the Ctrl, iKO, and idKO mice were fixed immediately and stained for Pax7. The mean number of the Pax7+ MuSCs/fiber was calculated.
  6. FACS‐isolated MuSCs were immediately fixed and stained for Pax7 (green) and DAPI (blue).
Data information: Scale bars: 50 μm. In (B, E), the data were presented as mean ± s.d. **P < 0.01; ***P < 0.001; unpaired Student's t‐test.