Notch signal interruption promotes CD4 and DP cell differentiation.

<p>CB and adult HSCs were grown in OP9Delta<sup>pos</sup> co-culture, then Notch signalling was removed for 7 days by transfer to OP9Delta<sup>neg</sup> co-culture or addition of DAPT. Cells were analysed by flow cytometry for CD4 and CD8 expression. (A) Representative dot plots for adult cells co-cultured for 70 days. (B–E) Adult (B, D) or CB (C, E) HSCs were grown in OP9Delta<sup>pos</sup> co-culture (▪) and then either transferred to OP9Delta<sup>neg</sup> co-culture (○) or incubated with DAPT (▵) for 7 days. Data points show the percentage of CD4<sup>+</sup> ISP (B, C) or CD4<sup>+</sup> CD8<sup>+</sup> DP (D, E) cells, n≥3, mean+SEM, * <i>p</i><0.05 for DAPT vs OP9Delta<sup>pos</sup> (<i>t</i>-test), ** <i>p</i><0.05 for OP9Delta<sup>neg</sup> vs OP9Delta<sup>pos</sup> (<i>t</i>-test). (F) CD14 (▪) expression plotted against DP expression (▴) for adult cells in OP9Delta<sup>pos</sup> co-cultures after Notch signalling was removed for 7 days. (G) Cell numbers during DAPT treatment. Paired comparisons of six control and DAPT – treated cultures, showing all HSC-derived cells (left) and DP cells (right).</p>