Cytochrome c

Western blotting and immunohistochemistry (IHC) were performed as previously described [15 (link)]. Antibodies against DNAJC19, PI3Kp85a, p-AKT(T308), AKT, GAPDH, CDH2, MMP2, TWIST, MYC, Caspase 3 and p-mTOR were purchased from Abcam (Cambridge, MA, USA. The catalog is ab230187, ab191606, ab8933, ab18785, ab9485, ab18203, ab37150, ab50887, ab32072, ab32351 and ab109268, respectively), and antibodies against CDH1, ERK1/2, p-P38, P38, MMP9, Snail, mTOR, NFkB-p65, p-ERK1/2, VIM, p-NFkB-p65, p-β-Catenin, β-Catenin, Slug were bought from CST (Massachusetts, USA. The catalog is 14472S, #9107, #4631, #8690, 13667s, 3879s, #2972, #8242, #4376, 3932s, 3033s, 2009s, 9562s, #9585, respectively). And antibody against FN1 was purchased for R&D (Minnesota, USA. The catalog is MAB19182). The antibodies against PARP1, Cytochrome C were bought from ZEN BIO (Chengdu, China. The catalog is 385279, 200,758, respectively).

Proteins were extracted from the BMSCs of each group of mice or from in vitro samples. The cytoplasmic and mitochondrial proteins were extracted using the Cytoplasmic and Mitochondrial Protein Extraction kit (Beyotime Biotechnology), according to the manufacturer’s instructions. Immunoblotting was conducted as previously described.^{93 (link)} Primary antibodies against P21 (BD Pharmingen) at 1:500, VDR (Santa Cruz Biotechnology) at 1:500, SOD1 (Santa Cruz Biotechnology) at 1:1 000, SOD2 (Santa Cruz Biotechnology) at 1:1 000, cytochrome C (Zen-Bioscience) at 1:500, AIF (Zen-Bioscience) at 1:500, cleaved caspase-3 (Zen-Bioscience) at 1:500 and Vinculin (Zen-Bioscience) at 1:2 000 were used. Immunoreactive bands were visualized with enhanced chemiluminescence (ECL) (Epizyme Biotech) and analyzed with a ChemiDoc^TM MP imaging system (Bio-Rad).