Figure 5. GC responses on Day 8 in adoptively transferred mice with different precursor frequency.

1. Precursor frequency in CLK21, CLK09 and CLK19 adoptively transferred mice. 8‐week‐old CD45.1 mice were adoptively transferred with 5 × 10⁵, 1 × 10⁵ or 2 × 10⁴ CD45.2 B cells from CLK21, CLK09 or CLK19 KI mice 1 day before immunization with eOD‐GT8 60mer. 12 h post‐adopt transfer, splenocytes were isolated and detected by FACS. X‐axis represents the cell number of transferred CD45.2 B cells, y‐axis represents the precursor frequency which is defined as (Number of CD45.2 binders)/(Number of B cells) for three lines. Each circle represents each mouse in each group. n = 2–4 mice/group for CLK21 and CLK09; n = 3 mice/group for CLK19. Bars indicate mean ± SD from mice in each group.
2. CD45.2 frequency within GCs on Day 8 in CLK adoptively transferred mice with different precursor frequency. Adoptively transferred mice with different precursor frequency were immunized with eOD‐GT8 60mer, on Day 8, the splenocytes were isolated and the CD45.2 populations were detected by FACS with the marker SSL⁺B220⁺CD95⁺CD38⁻CD45.2⁺. Gated plots represent the CD45.2 frequency within GCs.
3. Quantification of the frequency of CD45.2 and CD45.2 binders. Upper graph shows the quantification of CD45.2 frequency among GC in three adoptively transferred mouse models after the immunization of eOD‐GT8 60mer. The graphs below show the frequency of GC CD45.2 binders. X‐axis represents the different precursor frequency group. Y‐axis represents the percentage of GC (upper) and GC CD45.2 B cells (lower), respectively. Each circle represents one mouse. Mice from two independently repeated experiments were analyzed, n = 5–10 mice/group. Bars indicate geometric mean and geometric SD from mice in each group. Significant differences were calculated with Student’s t‐test and shown as: P > 0.05, no statistical significance (ns), *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. All P value analyses were calculated by GraphPad Prism V8.0.