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Fig. 2. PIPLC inhibits cell-cycle withdrawal and myotube formation in rodent and avian myoblasts. C2C12 (A-C), Sol8 (D-F), L6 (G-I), and QM7 (J-L) myoblasts were grown under differentiation conditions in the presence and absence of PIPLC (5 U/ml) until extensive myotube formation was observed in untreated cells: 5 days for C2C12 and Sol8 cells, 7 days for QM7 cells, and 10 days for L6 cells. All cultures were then labeled with BrdU. (A,D,G,J) Detection of all nuclei with DAPI and nuclei of cycling cells by immunostaining for BrdU demonstrated an increase in cycling cells and a decrease in multinucleated myotubes (as determined by phase microscopy) in the presence of PIPLC. Representative data are shown (n=5). Bar, 50 µm. (B,E,H,K) Fusion index [number of nuclei in fused cells (
2 nuclei)/total nuclei] was decreased in cells treated with PIPLC, compared with untreated cells. Data shown are mean±s.e.m. (n=4; *significant difference, P<0.05). (C,F,I,L) Percent BrdU-positive cells was increased in cells treated with PIPLC, compared with untreated cells. Data shown are mean±s.e.m. (n=5; *significant difference, P<0.05).
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