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Figure 5. Wnt/b-catenin signaling is blocked in hsDkk1 froglets. (A) Experimental scheme for in situ hybridization. Upper: Froglet forelimbs were amputated (amp: blue square) and heat-shocked (hs: red circle) at 8 dpa, and their blastemas were excised and fixed (fix: black triangle) 16 h after the heatshock for in situ hybridization. Lower: Froglet forelimbs were amputated (amp: blue square) repeatedly heat-shocked (hs: red circles) every other day until 8 dpa, and their blastemas were excised and fixed (fix: black triangle) 16 h after the last heatshock for in situ hybridization. (B and C) In situ hybridization on sectioned samples of froglet blastemas that had been heat-shocked as shown in the upper scheme in (A). The sectioned samples were hybridized with the fgf-8 antisense probe. (D and E) In situ hybridization of sectioned samples of froglet blastemas that were repeatedly heat-shocked as shown in the lower scheme in (A). Sectioned samples were hybridized with the fgf-8 antisense probe. To guarantee correct comparisons of gene expression levels, wild-type (B or D) and hsDkk1GFP froglet (C or E) sections were treated in exactly the same way. Scale bar = 100 mm.
doi:10.1371/journal.pone.0021721.g005 |
