Biological microscope system

The fluorescence spectra were acquired on a SPEX Fluorolog 3-TCSPC spectrometer equipped with 1 cm path length cuvettes. The pH value was measured by PHS-3C Precision Ph/Mv Meter. The cell images were acquired on an Olympus Biological Microscope System. The 3 peptides, TAM-PEP (R4): TAM-RRRRNLWAAQRYGRELRRMSDKFVD, TAM-PEP (R6): TAM-RRRRRRNLWAAQRYG RELRRMSDKFVD and TAM-PEP (R8): TAM-RRRRRRRRNLWAAQRYGRELRRMSDKFVD, were synthesized by GL Biochem (Shanghai, China) Ltd. GO (XF-020, 50–200 nm) was purchased from XianFeng Nano Co. (Nanjing, China). The cDNA3.1-Bcl-xL plasmid was constructed by Genecreate Biological Engineering Co. (Wuhan, China) Tris-HCl, IPTG, PMSF, Anti-Bcl-xL antibody, Anti-ACTB antibody, HRP-conjugated Goat Anti-Rabbit IgG, Cell Counting Kit-8, Cytochrome C, BSA, RNase and lysozyme were purchased from Sangon Biotechnology Co. (Shanghai, China) BeyoECL Star and trypsin digestion solution were purchased from Beyotime Biotechnology Co. (Shanghai, China) Lipo ™ 2000 was purchased from Thermo Fisher Scientific Co.(Beijing, China)The deionized water was prepared on a Milli-Q water purification system and used throughout all experiments.

Gastrocnemius specimens were fixed for 24 h in 4% paraformaldehyde and dehydrated in ascending grades of alcohol. Specimens were embedded in paraffin, sliced longitudinally into 4 μm-thick sections and mounted on glass slides. Tissue sections were deparaffinized in xylene and rehydrated through descending alcohols. Then, sections were microwaved in citrate-based buffer for antigen retrieval as per conventional methods. Sections were incubated overnight at 4 °C with a rabbit anti-desmin monoclonal antibody at a 1000-fold dilution. The reaction was developed using anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody (1:500) for 30 min. Sections were incubated with diaminobenzidine (DAB) for 5 min for visualization, and counterstained with hematoxylin for 10 min. The architecture of gastrocnemius muscle as well as desmin expression were observed on an Olympus biological microscope system (Tokyo, Japan)