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Figure 2


Fig. 2. Effect of ectopic Wnt proteins on satellite-cell proliferation. (A,B) Quantification of satellite-cell proliferation after a 36 hour culture period in the presence of Wnt1, Wnt3a, Wnt4, Wnt5a and Wnt6. All data are given as averages ± s.e.m. (A) Average cluster size following culture period: Wnt1 (2.6±0.09), Wnt3a (2.3±0.21) and Wnt5a (2.4±0.24) treated cultures all gave a greater number of satellite-cell progeny per cluster than LacZ controls (1.35±0.03), #P<0.0001, P=0.0001 and §P=0.0002, n=834, n=463, n=66 and n=51 clusters for LacZ, Wnt1, Wnt3a and Wnt5a, respectively. Wnt4 (1.03±0.01) and Wnt6 (1.13±0.02) showed significantly smaller cluster sizes than LacZ control cultures, *P<0.0001 and **P<0.0001, n=231 and n=548 clusters for Wnt4 and Wnt6, respectively. (B) Average total number of satellite cells per myofibre following culture period: Wnt1 (12.83±0.82) and Wnt3a (12.25±0.75) treated cultures both gave a greater number of satellite-cell progeny per myofibre than LacZ controls (10.45±0.57), #P=0.028 and P<0.05, n=124 and n=78 myofibres for Wnt1 and Wnt3a, respectively). Wnt5a gave a higher number of progeny cells per myofibre than control (11.09±1.1); however, this was not statistically significant. Wnt4 (7.18±0.47) and Wnt6 (7.83±0.53) both showed a significantly lower number of satellite cells per myofibre than LacZ control cultures, *P<0.001 and **P=0.0015, n=32 and n=92 myofibres for Wnt4 and Wnt6, respectively. (C-H) Single fibres cultured for 36 hours in Wnt1 (C,F), Wnt6 (D,G) and control cells (E,H). Wnt1 resulted in Pax7-expressing clusters of multiple cells (green arrows). By contrast, Wnt6 rarely showed Pax7-positive clusters and typically gave only single cells (green arrow). Control cells gave rise to small clusters (green arrow). (I) Quantification of satellite-cell activation by Wnt proteins. No statistical difference was detected in the number of MyoD+ cells per myofibre following coculture in Wnt1, Wnt6 or LacZ control conditions for 2 hours. n=21, n=25 and n=50 myofibres for Wnt1, Wnt6 and LacZ cultures, respectively. P=0.44 for Wnt1 and P=0.2 for Wnt6 when compared with the control. All results are mean ± s.e.m.





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