RUNX1 expression is aberrantly elevated in Gata1s erythroid cells. (A) Boxplots depicting the normalized expression of selected megakaryocyte lineage genes as determined by RNA-seq. The red horizontal line indicates the mean normalized expression, the light-red box represents the 95% confidence interval for the mean, and the blue box represents ± 1 SD. (B) Western blot of GATA1/GATA1s, RUNX1, and HSC70 in WT and G1s erythroid cells. Cell lysates were extracted from E13.5 total fetal liver cells. Each lane represents a different embryo. HSC70 is provided as a loading control. Fetal liver cells were isolated from E12.5 embryos and stained with erythroid surface markers (CD71 and Ter119) or megakaryocyte surface markers (CD41 and CD42), then assessed for RUNX1 expression by intracellular flow cytometry. Data from total fetal liver cells (C), individual stages of erythropoiesis (R0-R4/5) (D), and megakaryocytes (CD41−CD42− > CD41+CD42− > CD41+CD42+) (E) are shown. (F) Tracks of CUT&RUN-seq data corresponding to GATA1, GATA1s, and H3K27me3 as well as ATAC-seq at the Runx1 locus. Histograms were normalized to account for differences in the number of reads per library. The red dashed box (right) highlights the changes in recruitment of H3K27me3 and differential chromatin accessibility (ATAC-seq) at GATA1/GATA1s binding sites in the Runx1 proximal promoter.
Figure 5.

RUNX1 expression is aberrantly elevated in Gata1s erythroid cells. (A) Boxplots depicting the normalized expression of selected megakaryocyte lineage genes as determined by RNA-seq. The red horizontal line indicates the mean normalized expression, the light-red box represents the 95% confidence interval for the mean, and the blue box represents ± 1 SD. (B) Western blot of GATA1/GATA1s, RUNX1, and HSC70 in WT and G1s erythroid cells. Cell lysates were extracted from E13.5 total fetal liver cells. Each lane represents a different embryo. HSC70 is provided as a loading control. Fetal liver cells were isolated from E12.5 embryos and stained with erythroid surface markers (CD71 and Ter119) or megakaryocyte surface markers (CD41 and CD42), then assessed for RUNX1 expression by intracellular flow cytometry. Data from total fetal liver cells (C), individual stages of erythropoiesis (R0-R4/5) (D), and megakaryocytes (CD41−CD42− > CD41+CD42− > CD41+CD42+) (E) are shown. (F) Tracks of CUT&RUN-seq data corresponding to GATA1, GATA1s, and H3K27me3 as well as ATAC-seq at the Runx1 locus. Histograms were normalized to account for differences in the number of reads per library. The red dashed box (right) highlights the changes in recruitment of H3K27me3 and differential chromatin accessibility (ATAC-seq) at GATA1/GATA1s binding sites in the Runx1 proximal promoter.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal