Anti mouse ly 6a e sca 1 pe antibodies

To quantify the percentage of hematopoietic stem cells (HSCs), BM cells (1 × 10⁵ cells) were stained with anti-mouse CD117 (c-Kit) FITC and anti-mouse Ly-6A/E (Sca-1) PE antibodies (eBioscience, San Diego, CA, USA). The percentages of CD3⁺CD4⁺ (helper T cell, Th) and CD3⁺CD8⁺ (cytotoxic T cell, CTL) lymphocytes were also measured. After 12 h or 24 h of treatment, BM cells were harvested and stained with anti-mouse CD3 PE-Cyanine7, CD4 PE-Cyanine5 and CD8a PE antibodies (eBioscience, San Diego, CA, USA). The percentages of staining cells were counted and analyzed by FACS Calibur cytometer and CellQuest software (Beckman Coulter, Brea, CA, USA). Each experiment was performed in triplicate with three replicates each.

BMCs were obtained by femoral cavity flushing and filtered through a 200-eye cell sieve mesh to obtain a single-cell suspension. To quantify the percentage of HSCs, CD117- and sac-1-positive cells were washed and stained with anti-mouse CD117 (c-Kit) FITC and anti-mouse Ly-6A/E (Sca-1) PE antibodies (eBioscience). For the detection of apoptosis in BMCs, an Annexin V-FITC Apoptosis Detection Kit (eBioscience) was used. BMCs were stained with FITC-conjugated annexin V and propidium iodide (PI). Flow cytometry was performed using a fluorescence-activated cell sorter Calibur cytometer and analyzed with CellQuest software (Beckman Coulter, Brea, CA, USA).