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Figure 5


Fig. 5. The kinetics of blocking SHH signal transduction in cultured wild-type forelimb buds (E10.5, 34-36 somites) using 10 µM cyclopamine. (A) Loss of Gli1 transcription within 9 to 15 hours. T0: Gli1 expression in a non-cultured control limb bud. Wt: Gli1 expression in a wild-type limb bud cultured for 15 hours as a control. Cyc (9 hours): Gli1 expression in a wild-type limb bud cultured for 9 hours in the presence of cyclopamine. Cyc (15 hours): Gli1 expression in a wild-type limb bud cultured for 15 hours in the presence of cyclopamine. Complete loss of Gli1 expression occurs. Therefore, all subsequent experiments were carried out by culturing limb bud as in the presence of 10 µM cyclopamine for 15 hours. (B) Fgf4 expression in the AER is lost by culturing limb buds in the presence of cyclopamine for 15 hours (Cyc panel). (C) By contrast, Fgf8 remains expressed in the AER of cyclopamine treated limb buds (Cyc panel). All limb buds are oriented with anterior towards the top and posterior towards the bottom.





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