R26NotchIC Cells Undergo Accelerated Neural Specification

<div><p>R26NotchIC ES cells or parental control 46C cells cultured under monolayer differentiation conditions.</p> <p>(A) Typical FACS profile of <i>Sox1</i>GFP expression after 48 h. <i>Sox1</i>GFP+ cells were scored using gate M1. </p> <p>(B) Proportion of <i>Sox1</i>GFP+ cells at various time points (average of triplicates). </p> <p>(C–H) Intact cultures at 72 h of monolayer differentiation, shown in phase contrast or stained for markers as indicated.</p> <p>(I) Typical FACS profile of <i>Sox1</i>GFP expression after 5 d. M1 is the gate used throughout to quantify the proportion <i>Sox1</i>GFP+ cells: note that this FACS profile indicates a more striking difference between control and NotchIC populations than is reflected by our conservative quantification. </p> <p>(J and K) Quantitative RT-PCR for Oct4 and for FGF5 during monolayer differentiation of R26NotchIC cells (NotchIC) or control parental 46C cells. Levels were normalised to GAPDH and are displayed relative to expression in 46C ES cells.</p> <p>(L and M) FACS analysis of the proportion of <i>Sox1</i>GFP+ cells for R26-NotchIC cells or parental 46C cells at various time points from triplicate cultures at normal density (10⁴ cells/cm²) or higher density (3 × 10⁴ cells/cm²) (average of triplicates). </p>