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( A ) Flow cytometry analysis of control and Mir195 -expressing Lin – cells. HPCs from fetal livers of wild-type mice were cultured for 7 days on OP9 with SCF, <t>Flt3-ligand,</t> and IL-7, after infection with control or Mir195 retrovirus. Representative result of control (upper panel) and Mir195 (lower panel) viral infections is shown (n=3). ( B ) Outline of the in vitro culture system of Ebf1 -/- HPCs. ( C ) Flow cytometry analysis of control and Mir195 -expressing Ebf1 -/- HPCs. Shown data is representative of n=3. ( D ) Microarray analysis of Mir195 -expressing Ebf1 -/- HPCs. Log 2 fold-changes in the expression levels of genes related to B (left panel), T (middle-upper panel), NK (middle-lower panel), and myeloid (right panel) cell lineages were classified and are shown as colored columns. The analysis was carried out in duplicates.
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( A ) Flow cytometry analysis of control and Mir195 -expressing Lin – cells. HPCs from fetal livers of wild-type mice were cultured for 7 days on OP9 with SCF, Flt3-ligand, and IL-7, after infection with control or Mir195 retrovirus. Representative result of control (upper panel) and Mir195 (lower panel) viral infections is shown (n=3). ( B ) Outline of the in vitro culture system of Ebf1 -/- HPCs. ( C ) Flow cytometry analysis of control and Mir195 -expressing Ebf1 -/- HPCs. Shown data is representative of n=3. ( D ) Microarray analysis of Mir195 -expressing Ebf1 -/- HPCs. Log 2 fold-changes in the expression levels of genes related to B (left panel), T (middle-upper panel), NK (middle-lower panel), and myeloid (right panel) cell lineages were classified and are shown as colored columns. The analysis was carried out in duplicates.

Journal: eLife

Article Title: A single microRNA miR-195 rescues the arrested B cell development induced by EBF1 deficiency

doi: 10.7554/eLife.101510

Figure Lengend Snippet: ( A ) Flow cytometry analysis of control and Mir195 -expressing Lin – cells. HPCs from fetal livers of wild-type mice were cultured for 7 days on OP9 with SCF, Flt3-ligand, and IL-7, after infection with control or Mir195 retrovirus. Representative result of control (upper panel) and Mir195 (lower panel) viral infections is shown (n=3). ( B ) Outline of the in vitro culture system of Ebf1 -/- HPCs. ( C ) Flow cytometry analysis of control and Mir195 -expressing Ebf1 -/- HPCs. Shown data is representative of n=3. ( D ) Microarray analysis of Mir195 -expressing Ebf1 -/- HPCs. Log 2 fold-changes in the expression levels of genes related to B (left panel), T (middle-upper panel), NK (middle-lower panel), and myeloid (right panel) cell lineages were classified and are shown as colored columns. The analysis was carried out in duplicates.

Article Snippet: The infected and transduced Lin - cells were cultured and differentiated into B cells on OP9 cells in IMDM (Thermo Fisher) supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 0.1 mM non-essential amino acid solution, 50 μM 2-mercaptoethanol, 100 units/mL penicillin G, 100 μg/mL streptomycin (all from Wako), and 10 ng/mL recombinant SCF, IL-7, and Flt3-ligand (R&D Systems).

Techniques: Flow Cytometry, Control, Expressing, Cell Culture, Infection, In Vitro, Microarray