Figure 1. Mutant Ezh2 provides an advantage to activated B cells in expanding the GC reaction.

A. WT bone marrow (BM) (CD45.1) mixed with Ezh2^Y641F BM (CD45.2) was injected into Rag1^-/- mice, SRBC-immunized and euthanized 3, 8 or 20 days later.

B. Gating strategy of splenocytes of one representative sample.

C. Flow cytometry data of mice immunized for 8 days was analyzed by normalizing the percentage of CD45.1⁺ GC B cells (CD38^-FAS⁺) to their parental CD45.1⁺ B cells (B220⁺DAPI^-), and equivalent normalization with CD45.2⁺ populations. Each pair of connected dots represents a mouse (n=4); paired t tests.

D. Analysis of non-GC B cells (CD38⁺FAS^-) at day 8 was done as in C.

E. IF confocal microscopy images of chimeric splenic GCs at day 8 post-SRBC.

F. Quantification of PNA fluorescence was overlapped with CD45.1 and CD45.2 shown in E (and non-shown images), each pair of connected dots representing a single GC; paired t tests.

G-H. Analysis of GC B cells (G) and non-GC B cells (H) at days 3 and 20 post-SRBC were done as in C (n=4 per group).

I. IF confocal microscopy images of chimeric splenic GCs at day 3 post-SRBC.

J. Quantification of GCs shown in I (and non-shown images) was done as in F.

Results at day 8 are representative of 3 independent experiments.

See also Figure S1.