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Fig. 4. BMP2-mediated activation of a Wnt signal and Bmp2 expression. E14.5 rat cortical stem cells were plated at 5092 cells/cm2 and exposed to 20 ng/ml BMP2 for 0, 24 or 48 hours. (A) Activated ß-catenin levels increased after 24 and 48 hours, as shown by western blotting. Activated ß-catenin control-cell lysate and {alpha}-tubulin were included as references. (B-E) Immunocytochemistry, showing increased ß-catenin activation after 24 hours of BMP2 treatment; control cells show only few faintly positive cells; (B,D) DAPI staining indicates total cell nuclei. (F) RT-PCR time course analysis of medium-density cultures after exposure to FGF2 or FGF2 and BMP2. One day of BMP2 exposure was sufficient to upregulate transcription of endogenous Bmp2 mRNA. GAPDH expression is shown in reverse-transcribed (+) and non-transcribed (–) samples as loading controls. Bars, 20 µm.





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