Genome-wide reduction of transcription pausing upon granulocytic differentiation. (A) Metagene analysis showing transcriptionally engaged Pol II occupancy measured by GRO-seq on both sense and antisense strands in undifferentiated human CD34⁺ HSPCs (day 0) and 3-day differentiated cells (day 3). (B) Left, An illustration for calculation of PI by the ratio of Pol II occupancy at promoter vs gene body. The promoter is defined from 300 bp upstream to 300 bp downstream of the TSS. The gene body is defined from 300 bp downstream of the TSS until the end of the gene or up to 13 kb for genes longer than 13 kb. Right, Boxplot analysis to compare PI between HSPCs (day 0) and granulocytes (day 3). (C) Cumulative distribution function analysis to compare PI distribution in HSPCs (day 0) and granulocytes (day 3). (D) Genome browser captures of GRO-seq for granulocytic differentiation regulator genes (PU.1 and CSF3R) and progenitor genes (TAL1 and GATA2) in HSPCs (day 0) and granulocytes (day 3). Scale bar and gene diagram are depicted above the captures. Yellow shades highlight promoter peaks.