Fig. 3.

HMGB1 regulates myeloid differentiation. a qRT-PCR and b Western blot analyses of HMGB1 expression during PMA-induced macrophage-like differentiation of HL-60 and THP-1 cells and ATRA-induced granulocytic differentiation of HL-60 and NB4 cells. c Western blot analyses of HMGB1 expression during the in vitro macrophage-like and granulocytic differentiation of CD34⁺ HSPCs. d The expression of HMGB1 was analyzed by Western blot in THP-1 cells that were infected with lenti-shHMGB1 (or lenti-ctrl) followed by PMA induction for 48 h. e qRT-PCR detection of macrophage-like differentiation markers CD14, CD11b, and CSF1R in the infected and PMA-induced cells. Three independent experiments were performed, and data are means ± standard deviations. f CD14 expression was evaluated by cytometric analyses in infected and PMA-induced cells. Red and black curves show results for untreated cells and anti-CD14 antibody-stained cells, respectively. g The expression of HMGB1 was analyzed by Western blot in NB4 cells that were infected with lenti-shHMGB1 (or lenti-ctrl) followed by ATRA induction for 48 h. h qRT-PCR detection of granulocytic differentiation markers CD11b, MPO, and CSF3R in the infected and ATRA-induced cells. Three independent experiments were performed, and data are means ± standard deviations. i CD11b expression was evaluated by cytometric analyses in infected and ATRA-induced cells. Red and black curves show results for untreated cells and anti-CD11b antibody-stained cells, respectively. *P < 0.05 and **P < 0.01, Student’s t test.