Figure 4
From: PI3K induces B-cell development and regulates B cell identity
PI3K signaling regulates B cell commitment and plasticity. (a) Cells from bm-derived wt and p110dKO pre-B cell lines were cultured either in the presence of IL-7, M-CSF, or without cytokines and analyzed by flow cytometry. Numbers indicate percentages of cells in the respective regions at day 7 after beginning of treatment. (Data are representative of at least 3 independent experiments). (b) FACS-analysis of CD11b expression of measured in cells from bm-derived wt and p110dKO pre-B cell lines cultured in the presence of IL-7 (upper panel). p110dKO cells were cultured in IL-7 or M-CSF supplemented medium and CD11b expression was assessed by FACS analysis at day 8 after beginning of treatment (lower panel). (c) Cells from a bm-derived SLP-65-deficient pre-B cell line were treated with LY294002 (30 μM) or solvent (DMSO) for 16 h and subsequently cultured in medium supplemented with M-CSF. Csf1r and Notch1 mRNA levels were determined with specific primers by qRT-PCR using the SYBR-Green detection method (left panel). Results are shown as mean ± SD of 2 independent analyses, run as duplicates. Statistical significance was calculated using the Mann-Whitney Test or the t-Test. Cells from the respective culture conditions were further analyzed by flow cytometry (right panel).
