Figure 1.
Figure 1. Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).

Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).

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