Fig. 6

Analysis of BAZ1B knockout in HUDEP-2 cell lines. Relative mRNA expression in various cells from A humans and B mice. Data for both human and mouse mRNA expression were obtained from The Scripps Research Institute BioGPS database(46). C Western blot analysis of BAZ1B protein expression in WT (n = 3) HUDEP-2 cell cultures at D0, D4, and D7 of maturation. GAPDH was used as loading control. Quantification was performed using Image J. RPL: relative protein level. D Schematic of CRISPR/Cas9 designed used to generate BAZ1B KO HUDEP-2 cell lines. E Western blot analysis of BAZ1B protein expression in WT (n = 3) and BAZ1B KO (n = 3) HUDEP-2 cell lines. HSC70 was used as a loading control and quantification was performed using Image J. WT: wild type, KO: knock out. p-values were calculated with a two-tailed Student’s t-test. F Wright–Giemsa staining of WT and BAZ1B KO HUDEP-2 cells during D0, D4, and D7 of maturation. Arrows indicate BAZ1B KO cells that have not condensed their nuclei to the same degree as WT. G WT and BAZ1B KO cell expansion measured by Trypan exclusion. H Log2 values of fold-change for specific erythroid and hematopoietic genes at D0 and D6 of maturation. Green bar represents genes that typically are upregulated during erythropoiesis and the red bar represents genes that are typically downregulated during erythropoiesis. Error bars represent ± SEM in C &E and ± SD in A, B and G. Asterisks (*) indicate significance as follows: **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001