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. 2014 Dec 15;211(13):2617–2633. doi: 10.1084/jem.20140317

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© 2014 Di Foggia et al.

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Figure 4. — Conditional activation of Bmi1 expression in satellite cells does not improve skeletal muscle regeneration after traumatic injury in vivo. (a) Schematic temporal summary of the experiment. (b–d) Quantification of Bmi1 expression upon Tamoxifen injections in Pax7^Bmi1 mice compared with controls at RNA (b; mean ± SEM from two independent experiments with n = 5 controls and n = 3 Pax7^Bmi1; *, P < 0.05) and at protein level (via double immunolabeling for Pax7 and Bmi1; c; arrows indicate a Pax7^+ve cell that is included in the insert, representative results from one experiment with n = 3 controls and n = 3 Pax7^Bmi1); quantification of the average pixel intensity of Bmi1 staining in Pax7^+ve cells (d; mean ± SEM with n = 3 controls and n = 3 Pax7^Bmi1; ***, P < 0.001). (e) H&E staining of hind limbs 10 d.a.i. in injured controls and injured Pax7^Bmi1 mice (top, representative results from two independent experiments with n = 6 controls and n = 3 Pax7^Bmi1). Gomori Trichrome staining for endomysial fibrosis in the same samples of H&E (injured controls and injured Pax7^Bmi1 mice; bottom; representative pictures from one experiment with n = 3 controls and n = 3 Pax7^Bmi1). (f) Quantification of the ratio of regeneration in Pax7^Bmi1 mice compared with controls (f, mean ± SEM from two independent experiments with n = 6 controls and n = 3 Pax7^Bmi1). (g) Quantification of the CSA distribution of CNF in injured Pax7^Bmi1 and injured controls: the percentage of CNF of control mice (gray), Pax7^Bmi1 mice (blue), and the distribution of mature fibers (striped gray) is shown (mean ± SEM from two independent experiments and five independent experiments with n = 8 injured controls, n = 3 Pax7^Bmi1, and n = 15 uninjured controls; *, P < 0.05; **, P < 0.01; ***, P < 0.001). (h) Treadmill performance in terms of the time run (h, mean ± SEM from three independent experiments with n = 12 uninjured controls, n = 7 injured controls and n = 3 Pax7^Bmi1. **, P < 0.01) in injured control mice (gray) as compared with uninjured mice (striped), as well as in the distances run (i; mean ± SEM from three independent experiments with n = 7 injured controls, n = 3 Pax7^Bmi1, and n = 12 uninjured controls; *, P < 0.05; **, P < 0.01). Bars, 125 µm.