Sterile 1 pbs

Qishen Yiqi Drop Pill (lot number: 110708) was obtained from Tasly Pharmaceutical Co., Ltd. with 0.5 g per pouch. Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS) and sterile 1 × PBS were purchased from Gibco (Carlsbad). Primary antibodies including cleaved caspase‐3 polyclonal rabbit antibody, bcl‐2 polyclonal rabbit antibody, bax polyclonal rabbit antibody, p53 monoclonal mouse antibody and β‐actin monoclonal mouse antibody, as well as LY294002, the specific inhibitor of PI3K, were obtained from Abcam. DCFDA Cellular ROS Assay Kit was also purchased from Abcam. Horseradish peroxidase‐conjugated secondary antibodies were obtained from Invitrogen. ECL chemiluminescence detection kit was obtained from Amersham Pharmacia Biotech. RIPA lysis buffer, BCA protein assay kit, TUNEL kit and the in vitro toxicology assay kit based on 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide (MTT) were purchased from Sigma‐Aldrich. Polyvinylidene difluoride (PVDF) membranes were from Bio‐Rad. The MitoCapture mitochondrial apoptosis detection kit was obtained from BioVision. The CytoTox 96® non‐radioactive cytotoxicity assay kit was obtained from Promega. The goat anti‐rabbit Alexa Fluor 555 secondary antibody and 4,6‐Diamidino‐2‐phenylindole (DAPI) were obtained from Thermo Fisher Scientific.

CB was diluted 1:1 with sterile 1×PBS (Gibco by Life Technologies, California, USA) and MNCs were isolated via a density gradient centrifugation (Biocoll, 1.077 g/cm³, Biochrom Merck). For erythrocyte lysis, cells were resuspended in 5 ml of ice-cold ammonium chloride solution (pH = 7.4, University Clinic Düsseldorf) with three consecutive washing steps. MNCs were counted and further prepared for sorting. Monocytes, B cells, and T cells were depleted via MojoSort Streptavidin Nanobeads (BioLegend) using the supplier’s negative selection protocol as previously described (63 (link)). The cells were stained with a lineage panel as previously described (69 (link)), as well as CD94, CD127, CD117, and CD34 for sorting of CD34⁺ HPCs. Cell sorting was performed on a MoFlo XDP (Beckman Coulter).

CB was diluted 1:1 with sterile 1×PBS (Gibco by Life Technologies, California, USA) and MNCs were isolated via a density gradient centrifugation (Biocoll, 1.077 g/cm³, Biochrom Merck). For erythrocyte lysis, cells were resuspended in 5 ml of ice-cold ammonium chloride solution (pH = 7.4, University Clinic Düsseldorf) with three consecutive washing steps. MNCs were counted and further prepared for sorting. Monocytes, B cells, and T cells were depleted via MojoSort Streptavidin Nanobeads (BioLegend) using the supplier’s negative selection protocol as previously described (63 (link)). The cells were stained with a lineage panel as previously described (69 (link)), as well as CD94, CD127, CD117, and CD34 for sorting of CD34⁺ HPCs. Cell sorting was performed on a MoFlo XDP (Beckman Coulter).