The T490A mutation increases RAG2 colocalization with H3K4me3.

Pearson correlation values (ρ) for GFP-labeled RAG2 proteins and either H3K4me3 (A) or H3K9me3 (B). Pre-B cells expressing the indicated GFP-RAG2 were co-stained with separate antibodies to H3K4me3 and H3K9me3. Secondary antibodies labeled with either Alexa 647 or Cy3 were used to detect the H3K4me3 and H3K9me3 labeling, respectively. The columns labeled H3K4me3 are a positive control consisting of cells labeled with rabbit antibody to H3K4me3 and co-stained with Alexa488- and Cy3-labeled antibodies to the primary antibody. Each symbol represents measurement of an individual nucleus. Indicated are the mean (blue lines) and SEM (error bars), calculated from the mean of three or more trials, each trial consisting of measurement of at least 30 cells. The red lines indicate significance tests between corresponding double-labeled samples, comparing cells with PDA treatment to control cells without PDA.