Figure 2.
Reduced G1/S transition of AIDR112HiGB cells and GC B cells. (A) In vivo BrdU labeling of GC B cells on day 7 upon SRBC immunization. Schematics of experimental setup and representative FACS plots. Quantification: BrdU+ cells in GC: P = 0.0035; DZ: P = 0.0014; LZ: P = 0.0294, two-way ANOVA (n = 10–11, two experiments). Cell cycle distribution, G1: P = 0.0027; early S: P = 0.0002; two-way ANOVA (n = 5–6, one experiment). 7-AAD on a linear scale to reflect the amount of the DNA content. (B) EdU and BrdU double labeling in iGB cells. Schematics for experimental setup and representative FACS plots and quantification, BrdU single-positive cells at 4 h: ****P < 0.0001. Cell cycle distribution: G1, ****P < 0.0001; early S, ****P < 0.0001; late S, **P < 0.01; 7-AAD on a linear scale to reflect the amount of the DNA content (n = 4, two experiments). (C) iGB cell proliferation rate measured by dilution of CTV. Left: Histogram overlay of WT and AIDR112H cells on day 3 and day 4. Right: Quantification of the percentage of cells in each division. Value and error bar: mean ± SD. MFI, mean fluorescence intensity. Refer to the image caption for details. Panel A: The panel shows a combination of flow cytometry plots and bar graphs. The flow cytometry plots display BrdU incorporation in different cell populations (Non-Act FoB, GC, DZ, LZ) from WT and AIDR112H mice. The bar graphs quantify the percentage of BrdU positive cells and cell cycle distribution in GC B cells. The x-axis of the bar graphs represents different cell cycle phases (G1, early S, late S, G2 slash M), and the y-axis represents the percentage of cells. The data show significant differences in BrdU positive cells and cell cycle distribution between WT and AIDR112H mice. Panel B: The panel shows a combination of flow cytometry plots and bar graphs. The flow cytometry plots display EdU and BrdU double labeling in iGB cells at different time points (5 min, 1h, 2h, 4h). The bar graphs quantify the percentage of BrdU single positive cells and cell cycle distribution. The x-axis of the bar graphs represents different time points and cell cycle phases, and the y-axis represents the percentage of cells. The data shows significant differences in BrdU single positive cells and cell cycle distribution between WT and AIDR112H iGB cells. Panel C: The panel shows a combination of histogram overlays and line graphs. The histogram overlays display the proliferation rate of iGB cells measured by dilution of Cell Trace Violet (CTV) on day 3 and day 4. The line graphs quantify the percentage of cells in each division. The x-axis of the line graphs represents different cell divisions, and the y-axis represents the percentage of cells. The data shows the proliferation rate of WT and AIDR112H iGB cells over time.

Reduced G1/S transition of AID R112H iGB cells and GC B cells. (A) In vivo BrdU labeling of GC B cells on day 7 upon SRBC immunization. Schematics of experimental setup and representative FACS plots. Quantification: BrdU+ cells in GC: P = 0.0035; DZ: P = 0.0014; LZ: P = 0.0294, two-way ANOVA (n = 10–11, two experiments). Cell cycle distribution, G1: P = 0.0027; early S: P = 0.0002; two-way ANOVA (n = 5–6, one experiment). 7-AAD on a linear scale to reflect the amount of the DNA content. (B) EdU and BrdU double labeling in iGB cells. Schematics for experimental setup and representative FACS plots and quantification, BrdU single-positive cells at 4 h: ****P < 0.0001. Cell cycle distribution: G1, ****P < 0.0001; early S, ****P < 0.0001; late S, **P < 0.01; 7-AAD on a linear scale to reflect the amount of the DNA content (n = 4, two experiments). (C) iGB cell proliferation rate measured by dilution of CTV. Left: Histogram overlay of WT and AIDR112H cells on day 3 and day 4. Right: Quantification of the percentage of cells in each division. Value and error bar: mean ± SD. MFI, mean fluorescence intensity.

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