Internalization of ODN by hematopoietic progenitors derived from CD34⁺ cells. (A) TLR9 expression in pDCs, B cells, cDCs, and hematopoietic progenitors induced erythroid differentiation. CD34⁺ cells (day 0) were cultured in the liquid phase in erythroid medium with or without 4μM 2006-PO for 1 to 7 days. The relative gene expression levels of TLR9 were normalized with 28S transcripts. The inset represents amplification of the y-axis. The results are representative of 3 independent experiments. (B) ODN-rhodamine and early endosome expression. CD34⁺ cells were cultured in erythroid medium for 2 days and incubated with rhodamine-conjugated 2006-PO or 2243-PO. Results are representative of 3 independent experiments. (C-D) ODN-rhodamine and lysosome expression. CD34⁺ cells were cultured in erythroid medium for 2 days, incubated with LysoTracker Green and rhodamine-conjugated 2006-PO (C) or 2243-PO (D) and analyzed using confocal microscopy. Results are representative of 3 independent experiments.