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Fig. 7. Gene silencing β-catenin with siRNA enhances myogenic differentiation. (a) Western blotting analysis shows that transfection of β-catenin siRNA significantly reduced β-catenin levels in proliferating (GM) C2 cells compared to transfection with control siRNA. β-Catenin levels remained lower for at least 3 days following the switch to mitogen-poor medium (DM). (b,c) C2 cells transfected with (b) β-catenin siRNA showed enhanced myogenic differentiation compared to cells transfected with (c) control siRNA. (d,e) Immunostaining showed that satellite-cell-derived myoblasts also exhibited reduced β-catenin levels when transfected with (d) β-catenin siRNA compared with (e) control siRNA. (f,g) Silencing β-catenin in primary myoblasts later promoted differentiation and fusion into large myotubes after culture in (f) mitogen-poor medium compared with (g) control cells (quantified in h). (i) Reduced β-catenin levels also significantly decreased the number of (i) Pax7+ cells compared with (j) control cells (quantified in k). Nuclei were counterstained with DAPI (blue). (h,k) Quantification of experiments shown in f,g (h), and in i,j (k). Random fields were selected from three independent cultures and values expressed as the mean ± s.e.m.; *P<0.05, significantly different from control cultures.
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