Anti β actin polyclonal antibody

Loxl3^f/f and Col2a1–Loxl3^–/– mice were sacrificed via cervical dislocation, and their cochleae were dissected. Cochleae proteins were incubated in cell lysis buffer (P0013, Beyotime) and extracted using a homogenizer. Western blot analyses were performed as described previously (Hou et al., 2014 (link)). The following primary antibodies were used: anti-VEGFA polyclonal antibody (rabbit, 1:400, ABclonal) and anti-β-actin polyclonal antibody (rabbit, 1:3,000, Proteintech). All data are presented as mean ± standard error of the mean, and data analyses were performed using the Image J software. Student’s t test was used for single-factor experiments involving two groups. The significance level was set to p < 0.05 for all statistical analyses.

Recombinant omentin-1 protein was purchased from R&D Systems (Minneapolis, MI, USA). Antibodies for AMPKα, p-AMPKα (Thr172), Akt, p-Akt (Ser473), ACCα, p-ACCα were purchased from Santa Cruz Biotechnology Inc (Waltham, MA, USA). Runx2 or omentin antibodies were purchased from Abcam (Cambridge, UK). Anti-mouse and anti-rabbit monoclonal IgG peroxidase conjugate antibody, anti-β-actin polyclonal antibody were purchased from Proteintech (Rosemont, USA). AMPK inhibitor, Compound C, and inhibitor of Akt, LY 294002, were purchased from Calbiochem (San Diego, CA, USA). AMPK and Akt specific siRNA oligonucleotides as well as the control siRNA oligonucleotides were purchased from Ribobio (Guangzhou, China). The ALP assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The fetal bovine serum, penicillin, and ptreptomycin were purchased from Gibco-BRL Co., Ltd (Grand Island, NY, USA). Alizarin Red S, pbs and calcitriol were purchased from Sigma Chemical Co., Ltd (St.Louis, MO, USA).

NaHS was purchased from Sigma-Aldrich (St Louis, MO, USA), and the cigarettes were purchased from Guangdong Tobacco Industry Co., Ltd. (Guangzhou, China). SRT1720 and EX 527 were purchased from Selleck Chemicals (Houston, TX, USA). The TRIzol Reagent was purchased from Ambion (Life Technologies, CA, USA). The PrimeScript RT reagent Kit with gDNA Eraser was from Takara Bio Inc. (Takara, Shiga, Japan), and the SsoFast EvaGreen Supermix was obtained from Bio-Rad Laboratories, Inc. (CA, USA). The primary and second antibodies described in this study include: anti-Bcl-2 and anti-β-actin polyclonal antibodies were purchased from Proteintech (Chicago, IL, USA); anti-MFN1, anti-SIRT1, anti-FOXO3 and anti-Bax antibodies were purchased from Abclonal (Wuhan, China); anti-p21 and anti-p53 antibodies were purchased from Cell Signaling Technology (CA, USA); anti-FIS1 antibodies, and the horseradish peroxidase (HRP)-labeled Goat Anti-Rabbit/Mouse IgG (H+L) were purchased from Abcam Biotechnology (Cambridge, MA, USA). The poly-vinylidene fluoride (PVDF) membranes were from Millipore Corporation (Billerica, MA, USA). ECL-Plus detection kit probed was purchased from Tanon Science and Technology Co., Ltd. (Shanghai, China). Other reagents were all purchased from GBCBIO Technologies Inc. (Guangzhou, China) unless otherwise indicated.