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Fig. 4. OLP induction by SHH requires FGFR. (A) E13.5 mouse neocortical cells were cultured for 2 DIV in the presence or absence of FGF2 or SHHAg, with or without the FGFR inhibitor PD173074 or the Hedgehog inhibitor cyclopamine. The cultures were assayed for OLIG2 immunoreactivity at DIV2 or NG2 immunoreactivity at DIV4. Induction of both OLIG2-positive and NG2-positive cells by SHH was inhibited by PD173074. The inducing activity of FGF2 was unaffected by cyclopamine. (B) Cortical cells from mouse E13.5 embryos were cultured in the presence or absence of SHHAg or FGF2, PD173074, a combination of FGFR1IIIc and FGFR1ßIIIc extracellular domains (sFGFR1) or cyclopamine. The inducing effect of FGF2 was inhibited by both PD173074 and sFGFR1 but not by cyclopamine. The effect of SHH was inhibited by PD173074 but not by sFGFR1, suggesting that SHHAg activity requires ligand-independent activation of FGFR. (C) Dose-response curve showing inhibition of OLP induction by SHHAg in the presence of increasing concentrations of PD173074 at DIV4. Half-maximal inhibition occurs at 25 nM PD173074, as described for inhibition of FGFR1 itself (Dimitroff et al., 1999).

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