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. 2022 Apr 21;10:814250. doi: 10.3389/fcell.2022.814250

FIGURE 5.

FIGURE 5

Gene expression of limb patterning genes in stage 46 developing axolotl limb buds. (A) Expression of Frem3 in the distal epithelium. Some mesenchymal expression can be observed in the anterior/proximal mesenchyme highlighted with red arrowheads. (A′) Epithelial expression of Frem3 decreased and the anterior mesenchymal expression was significantly increased after C59 treatment shown by red arrowheads. (B) Epithelial expression of Mtrans with a scewing towards the posterior side of the limb. (B′) C59 treatment greatly decreased expression of Mtrans in the epithelium. (C) Bmp2 expression is expressed in the basal epithelium are indicated with orange arrowheads and mesenchymal expression is highlighted with red arrowheads. (C′) Bmp2 expression had the same expression domains after C59 treatment, highlighted by orange arrowheads (epithelium) and red arrowheads (mesenchyme). The blue star indicates autofluorescence. (D) Bmp7 expression in the epithelium indicated with orange arrowheads and mesenchymal expression in the distal and anterior mesenchyme indicated with red arrowheads. (D′) Bmp7 expression continued after C59 treatment with a decrease in the anterior mesenchyme. (E) Fgf10 expression throughout the mesenchyme and lower levels in the epithelium. (E′) Fgf10 expression was completely absent after C59 treatment. (F) Hand2 expression in the posterior limb mesenchyme. (F′) Hand2 expression decreased and expanded in anterior regions of the mesenchyme, highlighted by red arrowheads. (G) Gli3 expression mainly in the mesenchyme. (G′) Decreased Gli3 staining in the mesenchyme. Blue stars indicate non-specific staining. (H) Broad mesenchymal staining of Etv4 throughout the limb bud with a few positive spots in the epithelium, highlighted by red arrowheads. (H′) Decreased Etv4 expression after C59 treatment, but with the same expression pattern as (H). (I) Distal mesenchymal expression of Hoxa13. (I′) Lack of Hoxa13 expression after C59 treatment. The blue star indicates autofluorscent muscle. Scale bars in all panels are 50 µm.