Oxotremorine m oxo m

Oxotremorine-M (OXO-M) (Sigma-Aldrich) was dissolved in distilled water to make a 50 mM stock solution and further diluted in the bath solution for the final concentration 50 μM. BD1047 and BD1063 (Tocris Bioscience) were dissolved in distilled water to make a 50 mM stock solution and further diluted in the bath solution for the final concentrations. IVM (Sigma-Aldrich) was dissolved in DMSO to make a 10 mM stock solution and further diluted in the bath solution to 100 μM. ACh (Sigma-Aldrich) was dissolved in distilled water to make a stock solution and then diluted in the bath solution to 1 μM. Sources of other materials are described in the relevant methods.

sparc −/− mice were a gift from Dr. Neveen Said at University of Virginia, Charlottesville, Virginia, USA^{27 (link)} and were maintained in the accredited pathogen-free Second Xiangya hospital mice facility on a 12 h light/dark cycle^{28 (link)}. C57BL/6 mice were purchased from Model Animal Research Center of Nanjing University. All experiments were approved by the Animal Care Research Committee of Second Xiangya Hospital (ACRCSXH) and carried out as per ACRCSXH guidelines.
Human sparc cDNA clone was described by us before^{29 (link)} and subcloned into pShuttle vector (Clontech). Adenovirus expressing human SPARC was constructed using Adeno-X expression system (Clontech) as described before^{30 (link),31 (link)}. GAPDH, Horseradish peroxidase labeled donkey anti rabbit or donkey anti mouse antibodies were from Cell Signaling (Beverly, MA). Recombinant mouse SPARC protein (cat. number 942-SP-050) was purchased from R & D systems. Oxotremorine M (Oxo-M) and LY294002 were purchased from Sigma-Aldrich. CCG4986 was purchased from ChemBridge (San Diego, CA).