Cd38 clone hb7

Two patient-derived xenograft mice models were applied in this study to evaluate the impact of anlotinib on the regeneration capacity of CD34⁺CD38⁻LSCs and CD34⁺ LSCs respectively. The first patient-derived xenograft model was generated using CD34⁺CD38⁻LSCs sorted from bone marrow of an AML patient while the other one was built using bone marrow mononuclear cells from another patient with CD34⁺CD38⁺AML cells. CD34⁺CD38⁻LSCs or bone marrow mononuclear cells were pretreated with anlotinib (2.5 μM) or vehicle (PBS), respectively, for 12 h in vitro. To exclude the impact of apoptotic cells on the engraftment of LSCs, we detected percentages of apoptotic cells in each group. Then 1 × 10⁶ live cells were injected into the tail vein of NOD-Prkdc^−/−IL2rg^−/− (NSG) mice (DMO Ltd., Beijing, China) after 1 Gy irradiation. In order to confirm the engraftment, hCD45⁺ cells were detected in the peripheral blood. At the 4th week, the NSG mice were sacrificed to analyze the tumor burden with human CD45-FITC (clone HI30, Biolegend) and murine CD45-BV421 (clone 30-F11, Biolegend) in spleen and evaluate LSCs with CD34-APC antibodies (clone 581, Biolegend) and/or CD38 (clone HB7, Biolegend) in bone marrow using flow cytometry.