Figure 4
Muscarinic agonists reduce the expression of contractile phenotype marker smooth muscle myosin heavy chain (SM-MHC) in ASMCs. A. Western blot analysis with anti-SM-MHC and anti-β-actin antibodies in total protein extracts from ASMCs incubated for 15 days in serum free medium (control) or in medium containing 10% FBS, ACh (10-5 M) or CCh (3 × 10-7 M). B. Quantification of SM-MHC expression normalized to the expression of β-actin presented in A. C. Specific SM-MHC expression by indirect immunofluorescence in ASMCs incubated for 15 days in serum free medium (control) or in medium containing 10% FBS, ACh (10-5 M) or CCh (3 × 10-7 M). Nuclei were stained with DAPI (lower panel). D. Counting of the percentage of rabbit tracheal smooth muscle cells which express SM-MHC in C. Ν = 4 independent experiments. *P < 0.05 compared with control.