1. Loss of polarity gene Dlg1 leads to an expansion of C'-1 stage cells during pre-B cell differentiation.
Loss of polarity gene Dlg1 leads to an expansion of C'-1 stage cells during pre-B cell differentiation. A, experimental design of competitive bone marrow reconstitution. WT (CD45.1) and Dlg1-deficient (CD45.2) lin−c-kit+Sca-1+ cells were sorted by fluorescence-activated cell sorting (FACS) from the bone marrow and injected at a 1:1 ratio into lethally irradiated Rag-deficient recipients. B, competitive bone marrow reconstitution assay with Dlg1-deficient and WT cells. WT (CD45.1) and Dlg1-deficient (CD45.2) lin−c-kit+Sca-1+ cells were FACS-sorted from the bone marrow, mixed at a 1:1 ratio, reanalyzed, and injected i.v. into lethally irradiated Rag-1–deficient recipients. One month after injection, cells from the spleen and lymph nodes (LN) were analyzed by FACS for B220+ (B cells) or B220− (non-B cells) cell markers. Data shown are representative of two independent experiments (n = 4). C, quantification of the contribution of WT (CD45.1) or Dlg1-deficient (CD45.2) cells in the spleen and lymph nodes for both B and non-B cells. Data shown are the composite of two independent experiments (n = 4). D, representative flow cytometry analysis of Fr.C, C'-1, and C'-2 cells in Dlg1f/f CD19 Cre+/− c-MyceGFP/eGFP (knockout, KO) or Dlg1f/f Cre −/− c-MyceGFP/eGFP (WT) mice. E, quantification of percentage and (F) cell number (per 1 million bone marrow cells) of C'-1 and C'-2 cells in knockout vs. WT mice. Data shown are a composite of four independent experiments (n = 6 WT/n = 6 knockout).
Gabriel J. Sandoval et al. Cancer Immunol Res 2013;1:
©2013 by American Association for Cancer Research