Panel A shows box plots comparing bone marrow B-cell developmental populations and cell numbers between wildtype and Sorl1 knockout mice. Panel B shows box plots comparing splenic transitional, follicular, and marginal zone B-cell populations and cell numbers. Panel C shows box plots comparing B-cell and Cluster of Differentiation 4 positive T-cell frequencies and cell numbers. Panel D shows a schematic diagram of the hematopoietic stem cell transplantation workflow, including lentiviral transduction, immune reconstitution, and immunophenotyping with a representative flow cytometry plot. Panel E shows box plots comparing mCherry positive bone marrow B-cell developmental populations after hematopoietic stem cell transfer. Panel F shows box plots comparing mCherry positive splenic transitional, follicular, and marginal zone B-cell populations after hematopoietic stem cell transfer. Panel G shows box plots comparing mCherry positive B cells and Cluster of Differentiation 4 positive T cells after hematopoietic stem cell transfer.
B cell development is not affected by Sorl1 deletion. (A) Bone marrow B cell populations in WT and Sorl1−/− mice; top graph shows percentage, bottom shows cell counts. n = 8–11 mice from three independent experiments. (B) Spleen B cell populations in WT and Sorl1−/− mice; top graph shows percentage, bottom shows cell counts. n = 13 mice from three independent experiments. (C) Lymph node lymphocyte populations in WT and Sorl1−/− mice; top graph shows percentage, bottom shows cell counts. n = 18 mice from four independent experiments. (D) Schematic depicting setup and analysis of CRISPR-targeted bone marrow chimeras. (E) Development of B cell populations in chimeras’ bone marrow from Cd4- or Sorl1-targeted HSCs. % mCherry+ cells at each stage is normalized to the mean immature mCherry+ B cell population for each sgRNA per experiment. n = 15–22 mice from four independent experiments. (F) Development of B cell populations in chimeras’ spleen from Cd4- or Sorl1-targeted HSCs. % mCherry+ cells at each stage is normalized to the mean immature mCherry+ B cell population for each sgRNA per experiment. n = 15–22 mice from four independent experiments. (G) Development of B and T cell populations in chimeras’ lymph node from Cd4- or Sorl1-targeted HSCs. % mCherry+ cells at each stage is normalized to the mean immature mCherry+ B cell population for each sgRNA per experiment. n = 12–23 mice from four independent experiments. P values shown in A–C and E–G are calculated by two-way ANOVA. Data are shown as box and whisker plots, where the line indicates the median, the box represents the interquartile range, and whiskers denote the minimum and maximum values.