Hemogen /BRG1 cooperativity modulates promoter and enhancer activation during erythropoiesis [RNA-seq]

To gain insight into hemogen function during human erythropoiesis, RNA-seq was performed in different type of erythroid cells, such as WT and hemogen KD CD34+ cells, WT and hemogen KD HUDEP2 cells, WT and hemogen KO K562 cells. Notably, depletion of hemogen in these human erythroid cells significantly reduced the expression of genes associated with heme and hemoglobin synthesis, such as ALAS2, HMBS, GYPA, EPOR, and HBB, supporting a positive role of hemogen in erythroid maturation. To further explore hemogen function in mouse embryonic erythropoiesis, RNA-seq was performed in WT and hemogen KO E14.5 fetal liver cells and the data indicated hemogen play a consistently positive role in erythroid maturation-related genes as shown in the human cells, suggested hemogen is conserved activator during the mammalian erythropoiesis. Overall design: RNA-seq were performed using WT and KD hemogen CD34 cells at differentiation day 8, 10,12,14 and 16, and WT and KD HUDEP2 cells before differentiation (day0) and after differentiation day 3,5 and 7 to identify the key genes regulated by hemogen during erythrocyte maturation. RNA-seq was also perfromed in human K562 cells to further validate hemogen targets during the differentiation. RNA-seq in embryonic day E14.5 fetal liver cells between control and hemogen knockout mice to identify the differentially expressed genes which are essential for mouse erythropoiesis.