Igm pe cy5

Peripheral blood from each mouse was subjected to complete blood count. Tibias, femurs, iliac crests, spines, ulnae, radii, and humeri were harvested for bone marrow cells. Spleen and thymus were collected for lymphocyte staining. Flow cytometry staining for all hematopoietic subpopulations including T cell developmental stages was performed as previously described (Yu et al., 2015a (link)). For B cell development, the following scheme was used: B220-Pacific Blue (BD Biosciences 558108), IgM-PE-Cy5 (eBiosciences 15-5790-82), CD43-fluorescein isothiocyanate (FITC) (eBiosciences 11-0431-82), CD24-APC (Biolegend 101814), and BP1-PE (eBiosciences 12-5891-83). Definitions of stages of B cell maturation were as follows: A′ pre-pro-B (IgM⁻B220⁺CD43⁺BP1⁻CD24⁻), B′ pro-B (IgM⁻B220⁺CD43⁺BP1⁻CD24⁺), C′ pro-B (IgM⁻B220⁺CD43⁺BP1⁺CD24^lo), C″ pro-B (IgM⁻B220⁺CD43⁺BP1⁺CD24^hi), pro-B (IgM⁻B220⁺CD43⁺), pre-B (IgM⁻B220⁺CD43⁻), B progenitors (IgM⁻B220⁺), immature B (IgM⁺B220^lo), and mature B (IgM⁺B220^hi). For cell-cycle analysis, bromodeoxyuridine-FITC and 7AAD, or Ki67-FITC and DAPI staining were coupled with staining of specific populations to reveal their cell-cycle status.