Gene expression analyses of follicular and MZ B cells from normal and CD19:KLF3 mice. (A) Cluster analysis based on 358 genes identified here as differentially expressed in CD19:KLF3 follicular B cells versus normal follicular B cells (fdr < 0.05). As shown by the genealogic tree above, CD19:KLF3 follicular and normal MZ B cells are related. Indeed, the subset of CD19:KLF3 follicular B cells is more related to MZ B cells of normal or CD19:KLF3 mice than to normal follicular B cells, which appear as a distinct entity. (B) Venn diagram of genes identified by the 2 comparisons based on normal MZ B cells versus normal follicular B cells (black circle) or CD19:KLF3 follicular B cells versus normal follicular B cells (red circle) (fdr < 0.05). (C) Dot plot analysis for expression differences of 2737 genes identified here as being differentially expressed in normal MZ versus follicular B cells (x-axis) and their corresponding expression difference in CD19:KLF3 follicular B cells versus normal follicular B cells (y-axis; all dots). The 358 genes that were differentially expressed in CD19:KLF3 follicular and normal follicular B cells are shown in red. Note that in the arrays KLF3 is detected by a probe outside the coding region of KLF3 that is not part of the transgene. Thus, KLF3 appears here not to be expressed in transgenic mice, whereas indeed transgenic KLF3 is expressed efficiently. Indeed, transgenic KLF3 expression appears to limit the expression from the endogenous KLF3 locus (supplemental Figure 1A). Array experiments were performed twice with follicular and MZ B cells of normal and CD19:KLF3 mice (n = 4 each). Representative data of one of 2 experiments, giving similar results, are shown. (D) Western blot analysis for KLF2 among normal and CD19:KLF3 follicular B cells. Two KLF2 specific bands are revealed.32 Equal loading is demonstrated by reprobing for the ribosomal elongation initiation factor 4A (eIF4A). Note that the eIF4A signal appears in between the KLF2 specific bands revealed earlier. Also shown is the Ponceau S stain of the blot before probing. A second experiment was performed giving similar results.
