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Fig. 3. FGF2 is required for BMP2-mediated EMT. Analysis of p75^NGFR expression in E14.5 rat cortical explants cultured in basal medium (DMEM-F12 without N2) alone or supplemented with growth factors (see Table 1). Treatments include a 4-day first phase and a 5-day second phase. Low-magnification images show p75^NGFR cytoplasmic staining (green) with DAPI⁺ nuclei (blue); original explants are shown on the right edge of each image. Only media containing FGF2+BMP2 (I-L, P) were able to induce neural crest precursors as measured by p75^NGFR expression and migration from explant. Neither insulin nor IGF1 was required (I-K) and IGF was not sufficient (B,D,G) to induce p75^NGFR+ precursors. (M,N) Substitution of EGF for FGF2 was not sufficient for neural crest induction. (O,P) A small number of p75^NGFR+ cells with thin processes in three of 42 IGF1/IGF1+BMP2 treated explants (O) that differ from the flattened morphologies of FGF2+BMP2/FGF2+BMP2 treated explants (P). (Q) RT-PCR panel showing Snail2 induction only in explants cultured with both FGF2+BMP2. Abbreviations: F, FGF2; I, IGF1; E, EGF; B, BMP2. Factor concentrations listed in Materials and Methods. Bars, 80 µm (A-M); 80 µM (N-P).

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