Fig. 1. Development of adipocytes from ES cell-derived neuroepithelial precursors. Sox2-ßgeo/oct4-tk genetically engineered ES cells were treated with RA and then selected between day 6 (D6) and day 10 (D10) to enrich for neuroepithelial cells and to eliminate residual undifferentiated ES cells. They were then induced to differentiate towards the adipocyte lineage. (A) At various times, the cells were processed for quantitative PCR analysis using Oct4, Sox1, Sox2, Sox9, Sox10, FoxD3 or GAPDH probes. Data were normalised relative to GAPDH amplification and the highest expression was defined as 100%. Similar results were obtained in two independent experiments. +sel, with selection; -sel, no selection. (B) After selection (D10), neuroepithelial precursors were stained with anti-FoxD3, anti-Sox9 or anti-Sox10 antibody (red) to identify NC-like cells, and with bisbenzimide to identify cell nuclei (blue). 14 days after induction of adipocyte differentiation (+), adipocytes were identified either using their characteristic morphology (C) or by RT-PCR to detect FABP4 mRNA (D). The results in D are shown for two independent experiments. Scale bar: 100 µm.