Generation of Vav1 promoter-driven Asxl1^Y588XTg mice. (A) Schematic depiction of the Flag-Asxl1^Y588X transgene. P1, P2, and P3 showed the location of the primer pairs used for genotyping and quantitative PCR (qPCR). HS, hypersensitive to DNase-I; pA, polyadenylation region; ss, splice sites. (B) Genotyping PCR using genomic DNA from WT and transgenic mice with 2 sets of primers: P1 (for both transgenic and endogenous Asxl1) and P2 (Asxl1^Y588XTg specific). (C) RT-PCR showing the expression level of both endogenous ASXL1 and transgenic mutant Asxl1^Y588X using primer set P3. I and II indicate mice lines. Glyceraldehyde-3-phosphate dehydrogenase was used as a control. Error bars represent mean ± standard error of the mean (SEM). mRNA, messenger RNA. (D) Western blots showing ASXL1^aa1-587 expression and endogenous ASXL1 expression levels in the spleen (left) and BM (right) cells of WT and Asxl1^Y588XTg mice. β-action was used as a loading control. (E) Total offspring number for WT and transgenic mice.