Figure 4.

PLAG1-S enforces a pro-HSC transcriptional state. (A) Loci annotations and distribution of PLAG1-S binding sites in the Lin^-CD34⁺ genome identified by CUT&RUN. (B) Enriched motifs among PLAG1-S genomic binding sites determined by HOMER indicating the % of PLAG1-S targets bound to the consensus and P value of the enrichment relative to genome-wide background occurrence of the consensus. (C) Volcano plot of differential gene expression in PLAG1-S overexpressing Lin⁻CD34⁺ cells. Red- or blue-colored genes are significantly changed by adjusted P value < .05 and green- and purple-colored genes are directly bound by PLAG1-S. (D) PLAG1-S overexpression and shPLAG1 transcriptomic alignment to DMAP signatures of hematopoietic compartments.⁴⁹ Numbers above or below the bars indicate the empirical P value determined based on the percentage of times for which the observed value (set of up- or downregulated genes) was as large or larger in that population than random values (equal number of randomly selected genes) based on 1000 trials. (E) Enrichment map of significantly enriched gene sets (FDR < 0.1) in PLAG1-S^OE Lin⁻CD34⁺ cells compared with control. Genes bound by PLAG1-S in Lin-CD34⁺ cells (CUT&RUN q-value cutoff of 0.05) are intersected to gene sets by Mann-Whitney U test (P < .05) and the width of green edges correlates with increasing statistical significance of the overlap. Node size reflects the number of genes in the gene set. (F) Forty-one of 46 gene sets from the “Establishment Protein Localization Translation” cluster that are overrepresented among PLAG1-S genomic binding sites (g:Profiler FDR < 0.1) and the list of bound leading-edge genes driving negative enrichments in this cluster. See also supplemental Figure 4. FDR, false discovery rate.