Figure 2.
Figure 2. Reactive FDCs do not originate from circulating progenitors. (A) Wnt-1Cre Ubow+/− mice were injected with an emulsion of CFA/OVA in their ears and rear footpads. 3 wk later, inflamed auricular and popliteal draining LNs were sectioned, stained for CD21/35 and IgD expression, and analyzed by confocal microscopy. Insets display high-magnification views of FDCs. Data are representative of 3 different experiments (2 mice per experiment, 4 analyzed LNs per mouse). (B) WT mice were joined surgically with naive CD21Cre Ubow+/− µMt mice to create parabiotic mice. 3 mo later, when full chimerism was achieved, the WT partner was injected with CFA/OVA in its rear footpads. 3 wk later, popliteal LNs of both mice were sectioned, stained for CD21/35 and IgD expression, and the presence of colored FDCs in their LNs was analyzed by confocal microscopy. The development of mature FDC networks in the µMt mouse combined to the appearance of dTomato+ leukocytes in the WT partner indicates successful chimerism. Data are representative of 3 different experiments (2 mice per experiment). (C) CD21Cre Ubow+/+ chimeras were injected with an emulsion of CFA/OVA in their ears and rear footpads. 3 wk later, inflamed draining LNs were sectioned, stained for CD21/35, and IgD expression and analyzed by confocal microscopy. (D) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Data are representative of 4 different experiments (2 mice per experiment, 4 LNs analyzed per mouse). Each dot represents one follicle. In A and D, a two-tailed Student’s t test was used to determine significance. n.s = nonsignificant. Bars, 25 µm.

Reactive FDCs do not originate from circulating progenitors. (A) Wnt-1Cre Ubow+/− mice were injected with an emulsion of CFA/OVA in their ears and rear footpads. 3 wk later, inflamed auricular and popliteal draining LNs were sectioned, stained for CD21/35 and IgD expression, and analyzed by confocal microscopy. Insets display high-magnification views of FDCs. Data are representative of 3 different experiments (2 mice per experiment, 4 analyzed LNs per mouse). (B) WT mice were joined surgically with naive CD21Cre Ubow+/− µMt mice to create parabiotic mice. 3 mo later, when full chimerism was achieved, the WT partner was injected with CFA/OVA in its rear footpads. 3 wk later, popliteal LNs of both mice were sectioned, stained for CD21/35 and IgD expression, and the presence of colored FDCs in their LNs was analyzed by confocal microscopy. The development of mature FDC networks in the µMt mouse combined to the appearance of dTomato+ leukocytes in the WT partner indicates successful chimerism. Data are representative of 3 different experiments (2 mice per experiment). (C) CD21Cre Ubow+/+ chimeras were injected with an emulsion of CFA/OVA in their ears and rear footpads. 3 wk later, inflamed draining LNs were sectioned, stained for CD21/35, and IgD expression and analyzed by confocal microscopy. (D) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Data are representative of 4 different experiments (2 mice per experiment, 4 LNs analyzed per mouse). Each dot represents one follicle. In A and D, a two-tailed Student’s t test was used to determine significance. n.s = nonsignificant. Bars, 25 µm.

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