Anti heat shock protein 70 hsp70

Rat PCC PC12 cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% horse serum, 5% fetal bovine serum (FBS; Sigma-Aldrich, St Louis, MO, USA), and antibiotic/antimycotic at 37°C with 5% CO₂. The HSP90 inhibitor, NVP-AUY922, and 17-AAG were purchased from Selleck Chemicals (Houston, TX, USA) and solubilized in dimethyl sulfoxide (DMSO) to a concentration of 1 mM/mL before use. Cell Counting Kit-8 (CCK-8) was purchased from Dojindo (Tokyo, Japan). All primary antibodies (anti-β-actin, anti-cyclin B1, anti-Bax, anti-heat-shock protein 70 (HSP70), anti-phospho-AKT (S473), anti-AKT, anti-phospho-ERK1/2 (T202/Y204), anti-ERK1/2, anti-phospho-MEK, and anti-MEK) were acquired from Cell Signaling Technology (Boston, MA, USA).

Collagenase (type D), dispase, CaCl_2, and anti-myogenic factor 5 (MYF-5) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Anti-heat shock protein-70 (HSP-70) and anti-CD31 (PECAM-1) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-desmin, anti-Pax7, anti-laminin, anti-alpha smooth muscle actin (SMA), and chemiluminescent horseradish peroxidase (HRP) substrate were purchased from Abcam (Cambridge, MA, USA). Unless stated otherwise, all media and reagents were purchased from Thermo–Fisher Scientific (Waltham, MA, USA).