Fig. 7: Nfkbie deficiency induces germinal center generation and amplification in vitro.
a Scheme of the germinal center B cells (iGCB) and plasmablasts (iPB) in vitro culture system. b Total number of live Nfkbie+/+ and Nfkbie−/− cells cultured on 40LB with IL4 and analyzed on day 4 (D4). Each symbol represents one mouse (n = 3). c Flow cytometry plots show live iGCB cells (CD19+GL7+FAS+) and iPB (CD19+CD138+) generated from FoB and MZB cells. Histograms present percentages (upper) and absolute cell numbers (lower) of iGCB cells and iPB on day 4. Each symbol represents one mouse (n = 3). d Representative cell cycle profiles of iGCB cells (left) and iPB cells (right), labeled with Dapi and anti-Ki67, and analyzed by flow cytometry at day 4 (D4). Numbers indicate percentages of cells in the gated populations. Histograms show percentages of the iGCB cells (left) and iPB (right) in the S and G2/M phases of the cell cycle on day 4. Each symbol represents one mouse (n = 3). e Flow cytometry plots show representative apoptosis analysis of iGCB cells (left) and iPB cells (right), labeled with Annexin V and sytox, and analyzed by flow cytometry at day 4 (D4). Numbers indicate percentages of cells in the gated populations. Histograms show percentages of viable (defined as Annexin V− and sytox−) iGCB cells (left) and iPB (right) on day 4. Each symbol represents one mouse (n = 3). Data are mean ± SEM. *p < 0.05, **p < 0.01; ns, not significant.
