Modulation of miR-125b and miR-93 activity during NSC differentiation.

<p>(<b>A</b>) Representative GFP/mCherry expression in <i>stem/precursors</i> transduced with bdLV.miRT93 or bdLV.miRT125b (left), and their differentiated progeny (right). Percentages indicate GFP⁺ cells. MFI, mean fluorescence intensity of GFP signal. (<b>B</b>) Mean GFP fold repression (FR) measured in bdLV.miRT125b- and bdLV.miRT93-transduced cells as compared to bdLV.CTRL-transduced cells (mean ±SEM; n = 2 independent experiments). Calculations were done as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067411#pone-0067411-g002" target="_blank">Figure 2D</a>. (<b>C</b>) Co-expression of GFP and mCherry in bdLV.CTRL-transduced cells. Red, anti-mCherry antibody; green, GFP direct fluorescence. (<b>D</b>) Downregulation of GFP expression indicates increased activity of miR-125b at late stages of NSC differentiation in bdLV.miRT125b-transduced cells. (<b>E</b>) Faint GFP expression in bdLV.miRT93-transduced NSC cultures indicates robust activity of miR-93 in stem/precursor cells that is maintained in ≈50% of cells in progenitors and differentiated cells. Prec, <i>stem/precursors</i>; Prog, <i>progenitors</i>; Diff, <i>differentiated cells</i>; d, days in vitro. Data are expressed as percentage of GFP⁺ cells on transduced cells (mCherry⁺). Gray bars represent percentages in bdLV.CTRL-transduced cells. Data are the mean±SEM; n = 2 experiments, 3–5 coverlips/experiment. One-way analysis of variance followed by Bonferroni’s posttest. * p<0.05, *** p<0.001 versus bdLV.CTRL; § p<0.05 versus <i>stem/precursors.</i>