Figure 1.
NFIA-ETO2 blocks in vitro terminal differentiation of MEL and primary murine EBs. (A-C) NFIA-ETO2 expression (compared with vector-transduced cells) slightly increased proliferation (A) and the number of benzidine-positive cells (B) and reduced Ter119 surface expression on MEL cells grown for 4 days in 2% DMSO (C). (D) NFIA-ETO2–expressing MEL cells expressed significantly lower Hbα1 and Hbβ1 mRNA level than vector control-transduced cells. (E-F) NFIA-ETO2–expressing BM-derived mouse EBs proliferated significantly faster than vector-transduced control cells (CTRL) over 6 days in MM (E) or in DM (F) (n = 4). (G) Representative images of Wright-Giemsa-stained cytospin preparations from NFIA-ETO2–expressing primary mouse BM-derived EBs (top) compared with vector-transduced control cells (bottom). The left panels show EBs in MM (day 0), the middle panel EBs after 2 days in DM, and the right panel EBs after 4 days in DM. The small insert shows a reddish cell pellet in CTRL cells, whereas the pellet of NFIA-ETO2–expressing cells appeared white. Images were recorded with a 60× objective using a Nikon-TI. Scale bar, 50 μm. (H) NFIA-ETO2–expressing, FL-derived (E14.5) EBs continued proliferating, whereas cells overexpressing ETO2 started to decrease after 5 weeks of culture in MM. NFIA- and CTRL vector (pMSCV-GFP)–expressing cells could not be expanded. Data represent 1 out of 2 independent experiments. (I-J) NFIA-ETO2–expressing BM-derived EBs expressed lower levels of Hbα1 mRNA (I) and Gypa mRNA (J) than CTRL cells grown in MM (n = 3) and 24 hours in DM (n = 5), as assessed via qRT-PCR. (K) CD71 and Ter119 surface expression (in % of cells) on NFIA-ETO2– and CTRL, BM-derived EBs after 6 days in DM (n = 4). Values are presented as individual points; bar graphs represent the mean value of independent biological replicates (n); error bars are standard error of the mean. Statistical significances in was tested with paired two-tailed t tests.

NFIA-ETO2 blocks in vitro terminal differentiation of MEL and primary murine EBs. (A-C) NFIA-ETO2 expression (compared with vector-transduced cells) slightly increased proliferation (A) and the number of benzidine-positive cells (B) and reduced Ter119 surface expression on MEL cells grown for 4 days in 2% DMSO (C). (D) NFIA-ETO2–expressing MEL cells expressed significantly lower Hbα1 and Hbβ1 mRNA level than vector control-transduced cells. (E-F) NFIA-ETO2–expressing BM-derived mouse EBs proliferated significantly faster than vector-transduced control cells (CTRL) over 6 days in MM (E) or in DM (F) (n = 4). (G) Representative images of Wright-Giemsa-stained cytospin preparations from NFIA-ETO2–expressing primary mouse BM-derived EBs (top) compared with vector-transduced control cells (bottom). The left panels show EBs in MM (day 0), the middle panel EBs after 2 days in DM, and the right panel EBs after 4 days in DM. The small insert shows a reddish cell pellet in CTRL cells, whereas the pellet of NFIA-ETO2–expressing cells appeared white. Images were recorded with a 60× objective using a Nikon-TI. Scale bar, 50 μm. (H) NFIA-ETO2–expressing, FL-derived (E14.5) EBs continued proliferating, whereas cells overexpressing ETO2 started to decrease after 5 weeks of culture in MM. NFIA- and CTRL vector (pMSCV-GFP)–expressing cells could not be expanded. Data represent 1 out of 2 independent experiments. (I-J) NFIA-ETO2–expressing BM-derived EBs expressed lower levels of Hbα1 mRNA (I) and Gypa mRNA (J) than CTRL cells grown in MM (n = 3) and 24 hours in DM (n = 5), as assessed via qRT-PCR. (K) CD71 and Ter119 surface expression (in % of cells) on NFIA-ETO2– and CTRL, BM-derived EBs after 6 days in DM (n = 4). Values are presented as individual points; bar graphs represent the mean value of independent biological replicates (n); error bars are standard error of the mean. Statistical significances in was tested with paired two-tailed t tests.

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