Figure 6: Phenotypic analysis and characterization of cell dynamics in NKX2-3-expressing B cells. | Nature Communications

Figure 6: Phenotypic analysis and characterization of cell dynamics in NKX2-3-expressing B cells.

From: Homeobox NKX2-3 promotes marginal-zone lymphomagenesis by activating B-cell receptor signalling and shaping lymphocyte dynamics

Figure 6

(a) Representative flow cytometry profiles (grey filled histogram) of forward side scatter (FSC) and the indicated molecules at the surface of WT B cells and NKX2-3 transgenic (Tg) B cells of the specified ages (Tg6, 6-month-old mice, Tg12, 12-month-old mice and Tg18, 18-month-old mice). Dashed black line, maximum expression levels in WT B cells. Dotted histogram, isotype control. (b) Migration frequencies of WT and transgenic B cells of the indicated ages in Boyden chambers in the absence or presence of CXCL12, CXCL13 or CCL21. Data are the mean±s.d. (n=4). Error bars represent standard deviation (s.d.) in all cases. (c) Frequency of adhesion to ICAM-1 (top) or VCAM-1 (bottom) containing artificial membranes of WT and transgenic B cells. Data are the mean±s.d. (n=2). (d) Cell polarization frequencies, indicating the fractions of non-motile and motile cells, of WT and transgenic B cells settled on ICAM-1-membranes in the absence (top) or presence of CXCL12 chemokine (bottom); data are the mean±s.d. (n=2). (e) Mean velocity values of motile WT and transgenic B cells; data are derived from two experiments and each dot is a single cell. (f) Representative wide-field images of WT B cells (top) and transgenic B cells (bottom) after 24 h of culture. (g) IF analysis of WT and transgenic spleens from 12- and 18-month-old mice: (left) Clustering of B cells in LFA-1+ compartments in ageing Eμ-NKX2-3 mice: in WT mice (top) LFA-1+ (green) cells are restricted to the red pulp (arrow), whereas the follicles stained with anti-IgM (red; Fo) are devoid of such cells. Scale bar, 200 μm. (h) Representative flow cytometry profiles (filled histogram) of the indicated molecules at the surface of human B cells from healthy donors (HD) and patients (P3, patient positive for NKX2-3 expression; P4, P5, P6, patients negative for NKX2-3 expression). Dotted histogram, isotype control in each case. (i) Migration index of human B cells from healthy donors (n=4), P3 (patient positive for NKX2-3) and patients negative for NKX2-3 expression (n=7) in response to gradients of CXCL12.

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